Biomedical subjects

J E Pintar

Publications and source records attributed to J E Pintar.

At least 37 records · Page 2Linked to original sources

PubMed1999

Mu-opioid receptor modulation of calcium channel current in periaqueductal grey neurons from C57B16/J mice and mutant mice lacking MOR-1.

1. The actions of opioid receptor agonists on the calcium channel currents (IBa) of acutely dissociated periaqueductal grey (PAG) neurons from C57B16/J mice and mutant mice lacking the first exon of the mu-opioid receptor (MOR-1) were examined using whole cell patch clamp techniques. These effects were compared with the GABA(B)-receptor agonist baclofen. 2. The endogenous opioid agonist methionine-enkephalin (met-enkephalin, pEC50 6.8, maximum inhibition 40%), the putative endogenous mu-opioid agonist endomorphin-1 (pEC50 6.2, maximum inhibition 35%) and the mu-opioid selective agonist DAMGO (Tyr-D-Ala-Gly-N-Me-Phe-Gly-ol enkephalin, pEC50 6.9, maximum inhibition 40%) inhibited IBa in 70% of mouse PAG neurons. The inhibition of IBa by each agonist was completely prevented by the mu-receptor antagonist CTAP (D-Phe-Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr-NH2). The delta-opioid receptor agonists DPDPE ([D-Pen2,5]enkephalin, 1 microM) and deltorphin II (1 microM), and the kappa-opioid receptor agonist U-69593 (1-10 microM), did not affect IBa in any cell tested. 3. The GABA(B) agonist baclofen inhibited IBa in all neurons (pEC50 5.9, maximum inhibition 42%). 4. In neurons from the MOR-1 deficient mice, the mu-opioid agonists met-enkephalin, DAMGO and endomorphin-1 did not inhibit IBa, whilst baclofen inhibited IBa in a manner indistinguishable from wild type mice. 5. A maximally effective concentration of endomorphin-1 (30 microM) partially (19%), but significantly (P<0.005), occluded the inhibition of IBa normally elicited by a maximally effective concentration of met-enkephalin (10 microM). 6. This study indicates that mu-opioid receptors, but not delta- or kappa-opioid receptors, modulate somatic calcium channel currents in mouse PAG neurons. The putative endogenous mu-agonist, endomorphin-1, was a partial agonist in mouse PAG neurons.

Animals↗

PubMed1999

Identification of mouse CPX-1, a novel member of the metallocarboxypeptidase gene family with highest similarity to CPX-2.

The recent finding that Cpe(fat)/Cpe(fat) mice, which lack carboxypeptidase E (CPE) activity because of a point mutation, are still capable of a reduced amount of neuroendocrine peptide processing suggested that additional carboxypeptidases (CPs) participate in this processing reaction. Searches for novel members of the CPE gene family led to the discovery of CPD, CPZ, AEBP1, and CPX-2. In the present report, we describe mouse CPX-1, another novel member of this gene family. Like AEBP1 and CPX-2, CPX-1 contains an N-terminal region of 160 amino acids with sequence similarity to the discoidin domain of a variety of proteins. The 410-residue CP-like domain of CPX-1 has 54% to 62% amino acid sequence identity with AEBP1 and CPX-2 and 33% to 49% amino acid identity with other members of the CPE subfamily. However, several active-site residues that are important for catalytic activity of other CPs are not conserved in CPX-1. Furthermore, CPX-1 expressed in either the baculovirus system or the mouse AtT-20 cell line does not cleave standard CP substrates. Northern blot analysis showed the highest levels of CPX-1 mRNA in testis and spleen and lower levels in salivary gland, brain, heart, lung, and kidney. In situ hybridization of CPX-1 mRNA in embryonic and fetal mouse tissue showed expression throughout the head and thorax, with abundance in primordial cartilage and skeletal structures. In the head, high levels of CPX-1 mRNA were associated with the nasal mesenchyme, primordial cartilage structures in the ear, and the meninges. In the thorax, CPX-1 mRNA was expressed in multiple developing skeletal structures, including chondrocytes and perichondrial cells of the rib, vertebral, and long-bone primordia. Taken together, these findings suggest that it is unlikely that CPX-1 functions in the processing of neuroendocrine peptides. Instead, CPX-1 may have a role in development, possibly mediating cell interactions via its discoidin domain.

Amino Acid Sequence↗

PubMed1999

Generation of antisera to mouse insulin-like growth factor binding proteins (IGFBP)-1 to -6: comparison of IGFBP protein and messenger ribonucleic acid localization in the mouse embryo.

The insulin-like growth factor (IGF) system is an important regulator of fetal growth and differentiation. IGF bioavailability is modulated by IGF binding proteins (IGFBPs). We have generated six different antisera, directed to synthetic peptide fragments of mouse IGFBP-1 through -6. The specificity of the produced antisera was demonstrated by enzyme-linked immunosorbent assay, Western blotting, and by immunohistochemistry on sections of mouse embryos of 13.5 days post coitum. Specificity for the IGFBP-2 through -6 antisera also was confirmed immunohistochemically in liver and lung of corresponding gene deletion (knock-out) mutant mice and wild-type litter mates. Immunohistochemistry and messenger RNA (mRNA) in situ hybridization on sections of mouse embryos of 13.5 days post coitum revealed tissue-specific expression patterns for the six IGFBPs. The only site of IGFBP-1 protein and mRNA production was the liver. IGFBP-2, -4, and -5 protein and mRNA were detected in various organs and tissues. IGFBP-3 and -6 protein and mRNA levels were low. In several tissues, such as lung, liver, kidney, and tongue, more than one IGFBP (protein and mRNA) could be detected. Differences between mRNA and protein localization were extensive for IGFBP-3, -5, and -6, suggesting that these IGFBPs are secreted and transported. These results confirm the different spatial localization of the IGFBPs, on the mRNA and protein level. The overlapping mRNA and protein localization for IGFBP-2 and -4, on the other hand, may indicate that these IGFBPs also function in an auto- or paracrine manner.

Amino Acid Sequence↗

PubMed1998

The PACAP ligand/receptor system regulates cerebral cortical neurogenesis.

The PACAP ligand/type I receptor system is expressed throughout the embryonic nervous system, suggesting roles in regulating neural patterning and neurogenesis. In the forebrain, precursors of the six-layered cerebral cortex cease dividing in a highly reproducible spatiotemporal sequence. The time of cell cycle exit in fact determines neuron laminar fate. Our studies indicate that PACAP signaling may elicit cortical precursor withdrawal from the cell cycle, antagonizing mitogenic stimulators. PACAP inhibited embryonic day 13.5 rat cortical precursor [3H]thymidine incorporation, decreasing the proportion of mitotic cells. PACAP promoted morphological and biochemical differentiation, indicating that PACAP-induced cell cycle withdrawal was accompanied by neuronal differentiation. In vivo, embryonic cortex contains PACAP. In culture, 85% of cells expressed PACAP while 64% exhibited receptor. Co-localization studies indicated that PACAP ligand and receptor were expressed by the mitotic precursors that divided in response to bFGF, suggesting that precursors integrate mitogenic and anti-mitogenic signals to determine the timing of cell cycle exit. The expression of PACAP ligand and receptor in precursors raised the possibility of autocrine function. Indeed, peptide antagonists increased proliferation, suggesting that the PACAP system is expressed to elicit cell cycle exit. During ontogeny, an inhibitory signal, such as PACAP, may be required to counter the stimulatory activity of mitogenic bFGF and IGFI whose expression during cortical neurogenesis is sustained. The dynamic interplay of positive and negative regulators would regulate the timing of cell cycle withdrawal, and thus neuronal phenotype and laminar position.

Animals↗

PubMed1998

Developmental expression of VGF mRNA in the prenatal and postnatal rat.

VGF is a developmentally regulated, secretory peptide precursor that is expressed by neurons and neuroendocrine cells and that has its transcription and secretion induced rapidly by neurotrophins and by depolarization. To gain insight into the possible functions and regulation of VGF in vivo, we have characterized the distribution of VGF mRNA in the developing rat nervous system. VGF expression was first detectable at embryonic day 11.5 in the primordia of cranial, sympathetic, and dorsal root ganglia, and its distribution expanded throughout development to include significant expression throughout the brain, spinal cord, and retina of the adult rat. The earliest expression of VGF, therefore, appeared in the peripheral nervous system as developing neurons settled in their designated ganglia. In many regions of the brain, VGF mRNA levels were found to be highest during periods when axonal outgrowth and synaptogenesis predominate. Areas of the central nervous system that contain predominantly dividing cells never displayed any VGF mRNA expression, nor did the vast majority of nonneural tissues.

Animals↗

PubMed1998

Developmental expression of the mu, kappa, and delta opioid receptor mRNAs in mouse.

To characterize further the establishment of the opioid system during prenatal mouse development, we have examined the spatial and temporal expression patterns of mu, kappa, and delta opioid receptor mRNAs and find that the expression patterns of these mRNAs are distinct at all ages. Within the embryo, kappa is the first opioid receptor expressed, with transcripts detected in the gut epithelium as early as embryonic day 9.5 (E9.5). By E10.5, mu receptor expression is first detected in the facial-vestibulocochlear preganglion complex, whereas delta receptor mRNA is first detected at E12.5 in several peripheral tissues, including the olfactory epithelium, heart, limb bud, and tooth. In the brain, both mu and kappa mRNAs are first detected at E11.5 in the basal ganglia and midbrain, respectively. During mid-gestation and late gestation, the expression of both mu and kappa receptors extends to other brain regions that exhibit high expression in the adult, including the medial habenula, hypothalamus, pons, and medulla for mu and the basal ganglia, thalamus, hypothalamus, raphe, and ventral tegmental area for kappa. Thus by E17.5, many aspects of the adult expression patterns of mu and kappa receptors already have been established. Compared with mu and kappa, delta receptor mRNA expression in the brain begins relatively late, and the expression levels remain very low even at E19.5. In contrast to its late appearance in the brain, however, delta is the first opioid receptor expressed in the dorsal root ganglion, at E12.5, before its expression in the spinal cord begins at E15.5. Mu receptor is the first opioid receptor expressed in the spinal cord, at E11.5. These results extend previous ligand-binding data to significantly earlier ages and suggest that early developmental events in both neural and non-neural tissues may be modulated by opioid receptors. Several examples of possible autocrine and paracrine loops of opioid peptide and receptor expression have been identified, suggesting a role for these local circuits in developmental processes.

Animals↗

PubMed1998

Neuroendocrine protein 7B2 is essential for proteolytic conversion and activation of proprotein convertase 2 in vivo.

The 7B2 protein is widely distributed in neural and endocrine tissues. Its biological function was found to be related to the processing enzyme proprotein convertase 2 (PC2), a mammalian subtilisin/kexin-like endoproteinase that cleaves at specific single or multiple basic amino-acid residues. In order to examine the proposed function of 7B2 on PC2 in in vivo models, we first compared the distribution of 7B2 and PC2 mRNAs in the rat brain. Expression of 7B2 mRNA was found to be pan-neuronal, but additionally, we observed 7B2 mRNA in ependymal cells and in the subcommissural organ. Although the expression of PC2 mRNA was exclusively neuronal, it was more restricted, sparing some regions expressing high levels of 7B2. This finding suggests that 7B2 has an additional function in non-PC2-expressing cells. No evidence of PC2-positive/7B2-negative cells could be obtained in the adult rat brain. However, in the developing rat brain (E17), such regions were easily observed, showing higher levels of pro-PC2 (75 kD). Similarly, in the animal model of insulin-induced hypoglycemic shock, where adrenomedullary 7B2 expression is decreased, the ratio of pro-PC2 to mature PC2 (75 kD:68 kD) was observed to be increased. Finally, the human neuroepithelioma SK-N-MCIXC expresses PC2 but not 7B2. Accordingly, only inactive pro-PC2 forms were observed: 75-kD intracellular and 71-kD extracellular. After stable transfection of SK-N-MCIXC cells with 27-kD pro-7B2, mature and active (68-kD) PC2 was secreted into the medium. Our data demonstrate a critical role of 7B2 in the proteolytic conversion and activation of PC2 in vivo.

Adrenal Glands↗

PubMed1998

Expression of opioid receptors and ligands in pregnant mouse uterus and placenta.

The endogenous opioid system has been implicated in the regulation of hormonal secretion, pain perception, and uterine contractility during pregnancy, but there is only limited information about the cellular location of opioid receptor and opioid peptide gene expression in the pregnant rodent uterus and placenta. In this study, we have used in situ hybridization to identify expression sites of mRNAs encoding the delta (delta), kappa (kappa), and mu ( micro) opioid receptors as well as the endogenous opioid peptide precursors proenkephalin (PENK), prodynorphin (PDYN), and proopiomelanocortin (POMC) in pregnant mouse uterus and placenta. Soon after implantation, all three opioid receptor genes as well as POMC and PENK, but not PDYN, were detected in the uterine environment. Each expressed gene exhibited a distinct expression pattern that was generally retained until late gestation. The delta receptor and POMC were coexpressed in the trophoblast giant cells, which remained the only cells of the placenta/uterus to express these two genes throughout gestation. Cells expressing kappa receptors were absent from the placenta but instead were found in the basal part of the decidualized uterine endometrium. While kappa and micro receptors were transiently expressed in the uterine myometrium (until embryonic day 8.5), substantial levels of PENK were continuously detected in this region until at least embryonic day 18. In addition, complementary expression of the micro receptor and PENK genes in the uterus was detected. Taken together, these results suggest multiple roles for the opioid receptors and opioid peptides in maternal adaptation to pregnancy and in supporting embryo growth.

Animals↗

PubMed1997

Embryonic and uterine expression patterns of peptidylglycine alpha-amidating monooxygenase transcripts suggest a widespread role for amidated peptides in development.

Posttranslational processing of peptide precursors frequently includes COOH-terminal amidation by the bifunctional enzyme peptidylglycine alpha-amidating monooxygenase (PAM). We examined the ontogeny of PAM gene expression using in situ hybridization and detected expression in the cardiogenic region beginning at embryonic day 9 (e9) and in decidualizing uterine endometrium and myometrial smooth muscle at even earlier postimplantation stages. PAM expression in the CNS at e10 was highest in the dorsal spinal cord and floor plate and exhibited complex patterning in several CNS regions, including the ventricular zone, over the next several days with PAM expression first detected in neurons at e13. High levels of PAM expression characterized several nonneural cell populations as well, including limb mesoderm and the mesenchyme immediately adjacent to nasal, maxillary, palatal, and dental epithelia during tissue fusion and remodeling. Since alternative splicing generates PAM transcripts encoding proproteins that are differentially localized and processed, we used probes that distinguish major subsets of PAM transcripts to determine that transcripts encoding integral membrane PAM isoforms predominate in most, if not all, PAM-expressing cell types throughout development. Further, transcripts that encode soluble and cleavable PAM isoforms are essentially absent from two CNS areas that are rich in transcripts encoding integral membrane, bifunctional PAM: the ependymal region of the spinal cord and the ventricular zone of the hippocampus. These results provide evidence for widespread expression and cell-type-specific alternative splicing of PAM during development and raise the possibility that region-specific amidation of PAM substrates contributes significantly to several developmental processes.

Animals↗

PubMed1997

Ontogeny of prohormone convertases in rat prenatal development.

It has been well established that peptide precursors usually undergo limited proteolysis at pairs or single basic amino acids during their biosynthetic process. This posttranslational modification paradigm is common for numerous membrane-spanning and secreted proteins, neuropeptides, and peptide hormones of physiological significance, in which endoproteolytic cleavage is invariably essential for the accurate biosynthesis and full activity of the mature products. Establishment of an effective peptide profile is dependent on not only the presence of peptide precursor, but also the presence and the enzymatic specificities of cleavage enzymes. We have, therefore, characterized the spatial and temporal patterns of six subtilisin-like serine endoproteases known to be involved in proprotein processing, including furin, PC1, PC2, PC4, PC5, and PACE4, in rat prenatal development and related the results to the expression patterns of several peptide precursors. We have observed largely distinct and sometimes complementary expression patterns of individual PCs in various embryonic structures, suggesting PCs may be functionally distinct in processing different sets of proprotein substrates in development. From these studies, numerous tentative enzyme-substrate relationships in various embryonic structures have been proposed and should encourage more studies to test the in vitro cleavage potentialities of individual PCs toward these precursors. In the future, knowledge gained from these studies, when combined with insights gained from in vivo perturbation and genetic ablation studies, should lead to final comprehensive understanding of specific precursors cleaved by specific enzymes at specific cleavage sites in known spatial and temporal expression patterns during development.

Animals↗

PubMed1997

Insulin-like growth factor binding protein gene expression in the pregnant rat uterus and placenta.

While the insulin-like growth factor (IGF) system plays a fundamental role in regulating embryonic and placental growth, the specific contributions of the six IGF binding proteins (IGFBPs 1-6) to these processes are not well understood. We here focus on IGFBP expression in the extraembryonic environment, which both supports and constrains embryonic growth, and have used in situ hybridization to determine sites of IGFBP mRNA synthesis in the pregnant rat uterus and placenta. We find that all IGFBPs are expressed in distinct, changing patterns in the uterine endometrium, at the decidual boundary, in the decidual vasculature, and in the myometrium during pregnancy. Within the endometrium, the most prominent change is that expression of IGFBP-1 begins in some, but not all, endometrial glands prior to implantation and then expands to include all secretory epithelia shortly after implantation. During the period of rapid decidual proliferation that follows implantation, IGFBP-3, -4, and -5 transcripts are all detected in a laminar array at the boundary between the decidua and the nondecidualized endometrium. In the decidual vasculature at Day (d) 8.0, both IGFBP-3 and IGFBP-4 mRNAs are detected in dilating blood vessels, with BP-3 most prominent in the antimesometrial plexus and BP-4 primarily at the mesometrial pole. Later (d11.5), all decidual vessels express high levels of IGFBP-3 and lower levels of IGFBP-4 mRNAs. Finally, changes in expression of several IGFBPs also occur within the myometrium during pregnancy. For example, IGFBP-2 is expressed in the inner circular layer shortly after implantation, and expression increases through late gestation. In contrast, IGFBP-5 hybridization occurs over both myometrial layers before implantation, but decreases in intensity and spatial distribution as pregnancy proceeds. Finally, and most strikingly, IGFBP-6 expression, barely detectable in the d7.0 myometrium, gradually increases until it is very strongly transcribed during the placental stages. Taken together, these observations suggest multiple roles for IGFBPs in supporting implantation, regulating the extent of decidualization, modulating local levels of vascular IGFs, and regulating uterine muscular growth.

Animals↗

PubMed1997

The developmental expression in the rat CNS and peripheral tissues of proteases PC5 and PACE4 mRNAs: comparison with other proprotein processing enzymes.

Many peptides modulating cellular growth and differentiation in development are first synthesized as precursors that require proteolytic processing by the "prohormone convertase" (PC) family of endoproteases. Using in situ hybridization, we have here determined that two recently identified PC members, PC5 and PACE4, are expressed prenatally in spatial and temporal patterns that are each unique and distinct from those of previously characterized PCs. PC5 mRNA is first detected at e9 in highly restricted regions of the neural tube, in caudal myotomes, and at the materno-embryonic junction of the uterus. At e10, restricted PC5 mRNA expression is detected in the optic and otic vesicles, the roof of midbrain, and trunk myotomes. By midgestation (e13-e16), PC5 mRNA expression in the developing nervous system has expanded to multiple regions including hippocampus, thalamus, hypothalamus, brain stem, and spinal cord. By midgestational stages, PACE4 mRNA is expressed in multiple regions of the developing nervous system, generally distinct from PC5, and including a uniquely high level of expression in the ventricular zone of the hippocampus. In several peripheral organ systems, including lung and gut, we observed remarkably complementary patterns of PC5 and PACE4 expression. In addition, PACE4 transcripts are expressed in the heart and liver, whereas PC5 is expressed in the adrenal and kidney primordia. These results suggest that both PC5 and PACE4 may be involved in neuropeptide precursor processing in the developing nervous system and peripheral tissues with the general nonoverlapping expression patterns suggesting that PC5 and PACE4 may process distinct sets of proprotein substrates.

Animals↗

PubMed1996

Ontogeny of basal and regulated secretion from POMC cells of the developing anterior lobe of the rat pituitary gland.

Proopiomelanocortin (POMC)-producing cells are present in both the anterior (AL) and intermediate (IL) lobes of the adult rat pituitary. Both cell types are derived from a single embryonic rudiment, Rathke's pouch, and synthesize the same hormone precursor, POMC, but differ in the pattern of precursor processing and regulation of peptide secretion. Here we have used the reverse hemolytic plaque assay to determine the ontogeny of basal and regulated secretion by AL POMC cells. Basal secretion of beta-endorphin was first observed at Embryonic Day 13.5 (e13.5), the age when POMC-derived peptides were first detected immunocytochemically. Peptide secretion stimulated by corticotropin releasing hormone (CRH; 10(-8) M) was first detected at e15.5. The CRH-stimulated secretion involved two different components: (1) an increase in the amount of hormone secreted per cell (increase in plaque size) as well as (2) an increase in the number of plaque-producing cells. The greatest stimulation by CRH was observed at e16.5, a time when AL POMC mRNA levels increase significantly and when CRH neurons are first detected immunocytochemically in the hypothalamus. CRH-stimulated secretion, but not basal release, was inhibited by preincubation with dexamethasone (DEX; 10(-6) M) as early as e15.5, while the dopamine agonist ergocryptine (ERG; 10(-6) M) did not alter basal or CRH-stimulated release at any age studied. These results demonstrate that AL POMC cells are capable of hormone secretion as early as POMC peptides are first detected immunocytochemically and that these cells respond in an adult-like manner to physiological regulators soon after their initial appearance. Moreover, these responses remain unaltered during the early postnatal stress-nonresponsive period, suggesting that deficits at this time must lie at a level other than the corticotroph. Taken together, these results show that AL POMC cells possess functional regulatory receptor systems prior to maturation of the hypothalamic-hypophyseal portal system.

Age Factors↗

PubMed1996

Ontogeny of basal and regulated proopiomelanocortin-derived peptide secretion from fetal and neonatal pituitary intermediate lobe cells: melanotrophs exhibit transient glucocorticoid responses during development.

Proopiomelanocortin (POMC)-producing cells comprise nearly 100% of the adult rat intermediate lobe (IL) hormone-producing cells. Secretion by these cells in the adult is primarily under negative regulation by dopamine. Although the POMC-derived peptide alpha-MSH has been detected in plasma of fetal rats, the secretory capability of fetal melanotrophs has not yet been examined directly. Here we have used the reverse hemolytic plaque assay to assess, at the single-cell level, basal and regulated release by melanotrophs from fetal and early postnatal ages. Basal secretion was detected at the earliest age examined [Embryonic Day 17.5 (e17.5)], but CRH (10(-8) M) stimulated secretion was not observed until e19.5. As development proceeded, CRH increased both individual plaque sizes and the percentage of melanotrophs stimulated to secrete. An unexpected, transient inhibition of CRH stimulated release from melanotrophs by dexamethasone (DEX, 10(-6) M) was observed from e19.5-p2 (postnatal Day 2). By p3, however, DEX no longer inhibited melanotroph secretion while inhibition of CRH-stimulated release from p3 corticotrophs was readily detected. The dopamine agonist ergocryptine (ERG, 10(-6) M) inhibited basal secretion from melanotrophs, but not corticotrophs, at all ages examined. Taken together, these results indicate that melanotrophs undergo a maturation process in which they are initially nonresponsive to CRH, next possess functional CRH and steroid receptors, and finally, undergo functional uncoupling of steroid receptors which characterizes the adult IL. The loss of steroid-mediated inhibition of stimulated secretion parallels the arrival of catecholaminergic input into the IL. In contrast, the early response of melanotrophs to dopaminergic agonists, which can be detected 1 week prior to arrival of catecholaminergic fibers into the neurointermediate lobe, appears to be an intrinsic feature of these cells that is never present in corticotrophs.

Age Factors↗

PubMed1996

Genetic approaches to the function of insulin-like growth factor-binding proteins during rodent development.

Gene targeting provides a direct method for introducing mutations into specific mouse loci. This approach has been used productively to demonstrate that insulin-like growth factor (IGF) peptides and receptors are required for normal prenatal growth. Six genes comprising a third major component of the IGF system, the IGF-binding proteins (IGFBPs), are all expressed during prenatal rodent development. One of these genes, IGFBP-2, has also been disrupted using gene targeting, and homozygous null BP-2 mice are characterized by a decreased spleen size and an increase in circulating levels of other IGFBPs. These alterations are less dramatic than initially expected based on the fetal IGFBP-2 expression pattern. These results are discussed in light of both other genetic ablations involving members of gene families and in the context of the expression of other IGFBPs in rodent fetal and uterine tissues.

Adaptation, Physiological↗

PubMed1995

Genetic ablation of IGFBP-2 suggests functional redundancy in the IGFBP family.

Gene targeting allows mutations to be introduced selectively into any mouse locus of interest. This approach has already been used to demonstrate that insulin-like growth factor (IGF) peptides and receptors are required in vivo for normal prenatal growth. One of the IGFBP genes, IGFBP-2, has also been disrupted using gene targeting, and homozgyous null BP-2 mice are characterized by a decreased spleen size most apparent during early postnatal stages and increased adult circulating levels of several other IGFBPs. These alterations are considered less dramatic than the phenotypes initially predicted based on the fetal IGFBP-2 expression pattern, although several physiological paradigms can be envisioned that will provide additional tests for specific aspects of IGFBP function. Since all six IGFBP genes are expressed during prenatal rodent development, as well as in adult tissues, the IGFBP-2 null phenotype must also be compared with genetic ablations involving members of other gene families and in the context of the other IGFBP expression patterns in rodent embryonic, extraembryonic, and uterine tissues.

Animals↗

PubMed1994

Expression of the gene for the neuronal intermediate filament protein alpha-internexin coincides with the onset of neuronal differentiation in the developing rat nervous system.

While neurofilaments have long been considered early markers of neuronal differentiation, they cannot be detected in most newly postmitotic neurons of the developing central nervous system (CNS). Here we show that these neurons already express the neuronal intermediate filament protein alpha-internexin at high levels. alpha-internexin is expressed by most, if not all, neurons as they begin differentiation and shows no overlap with vimentin, whose expression in the CNS is restricted to mitotic neuronal precursors. In the adult, alpha-internexin is the only intermediate filament gene expressed by the cerebellar granule cells, the source of the thin-caliber parallel fibers; conversely, neurofilament proteins are highly expressed in large neurons, which express alpha-internexin at low levels. These data suggest that neuronal intermediate filaments may regulate axonal stability and/or diameter through changes not only in their number, but also in their subunit composition.

Animals↗

PubMed1994

Distinct expression patterns of insulin-like growth factor binding proteins 2 and 5 during fetal and postnatal development.

Insulin-like growth factors (IGFs), when isolated from serum or tissue fluids, are usually found as part of a protein complex which also contains one of several IGF binding proteins (IGFBPs). Although some IGFBPs have been shown to alter interactions of IGFs with their receptors in vitro and can modify the responses of cultured cells to exogenous IGFs, the in vivo functions of IGFBPs remain unclear. This study examines expression of a recently described IGFBP gene, IGFBP-5, in the rat embryo and fetus and in selected adult tissues. Embryonic IGFBP-5 messenger RNA (mRNA) can be detected as early as embryonic day 10.5 and has an mRNA expression pattern distinct from the previously characterized pattern of IGFBP-2 mRNA expression. Major sites of IGFBP-5 expression during early postimplantation stages of development include the notochord, the floor plate, regions of the surface ectoderm, muscle precursor cells, and specific axial regions of neuroepithelium. Later in development IGFBP-5 mRNA is found in several regions of the central nervous system, including the proliferative zone of the external granule layer of the cerebellum and the mitral neurons of the olfactory bulb, as well as in muscle precursor populations of the developing limb, and in most cells of the anterior pituitary. In addition, only a subset of pituicytes in the adult posterior pituitary express IGFBP-5, which provides the first evidence that this cell population is biochemically heterogeneous. Taken together, these data suggest functions for IGFBP-5 during development of several organ systems.

Aging↗