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Biomedical subjects

R Jaenisch

Publications and source records attributed to R Jaenisch.

At least 109 records · Page 6Linked to original sources
PubMed1996

Prenatal and postnatal requirements of NT-3 for sympathetic neuroblast survival and innervation of specific targets.

Postnatal homozygous neurotrophin-3 mutant mice display a loss of about half the sympathetic superior cervical ganglion (SCG) neurons (Ernfors, P., Lee, K.-F., Kucera, J. and Jaenisch, R. (1994a) Cell 77, 503-512; Farinas, I., Jones, K. R., Backus, C., Wang, X. Y. and Reichardt, L. F. (1994) Nature 369, 658-661). We found that this loss is caused by excessive apoptosis of sympathetic neuroblasts leading to a failure to generate a normal number of neurons during neurogenesis. NT-3 was also found to be required postnatally. In Nt-3-/- mice, sympathetic fibers failed to invade pineal gland and external ear postnatally; whereas other targets of the external and internal carotid nerves, including the submandibular gland and the iris, displayed a normal complement of sympathetic innervation. Sympathetic fibers of mice carrying one functional copy of the Nt-3 gene (Nt-3+/- mice) invaded the pineal gland, but failed to branch and form a ground plexus. Cultured neonatal sympathetic neurons responded to NT-3 by neurite outgrowth and mRNA upregulation of the NT-3 receptor, trkC. Exogenously administered NT-3 promoted sympathetic growth and rescued the sympathetic target deficit of the mutant mice. We conclude that NT-3 is required for the survival of sympathetic neuroblasts during neurogenesis and for sympathetic innervation and branching in specific targets after birth.

Aging↗
PubMed1995

Dependence of developing group Ia afferents on neurotrophin-3.

At birth, group Ia proprioceptive afferents and muscle spindles, whose formation is Ia afferent-dependent, are absent in mice carrying a deletion in the gene for neurotrophin-3 (NT-3-/-). Whether Ia afferents contact myotubes, resulting in the formation of spindles which subsequently degenerate, or whether Ia afferents and spindles never form was examined in NT-3-/- mice at embryonic days (E) 10.5-18.5 by light and electron microscopy. Three sets of data indicate that Ia neurons do not develop and spindles do not form in NT-3-deficient mice. First, peripheral projections of Ia afferents did not innervate hindlimbs of NT-3-/- mice, as reflected by a deficiency of nerve fibers in limb peripheral nerves and an absence of afferent nerve-muscle contacts and spindles in the soleus muscle at E13.5-E18.5. Second, central projections of Ia afferents did not innervate the spinal cord in the absence of NT-3, as shown by an atrophy of the dorsal spinal roots and absence of afferent projections from limb musculature to spinal motor neurons at E13.5 or E15.5. Lastly, the lumbar dorsal root ganglia (DRGs) at E10.5-E14.5, the stages of development that precede or coincide with the innervation of the spinal cord and hindlimbs by Ia afferents, were 20-64% smaller in mutant than in wild-type mice, presumably because the cell bodies of Ia neurons were absent in embryos lacking NT-3. The failure of Ia neurons to differentiate and/or survive and Ia afferent projections to form in early fetal mice lacking NT-3 suggests that NT-3 may regulate neuronal numbers by mechanisms operating prior to neurite outgrowth to target innervation fields. Thus, developing Ia neurons may be dependent on NT-3 intrinsic to the DRGs before they reach a stage of potential dependence on NT-3 retrogradely derived from skeletal muscles or spinal motor neurons.

Animals↗
PubMed1995

Generation of committed erythroid BFU-E and CFU-E progenitors does not require erythropoietin or the erythropoietin receptor.

Erythropoietin (EPO) is the principal growth factor regulating the production of circulating erythrocytes. We introduced null mutations into both Epo and the EPO receptor (EpoR) gene. Both heterozygotes appeared normal. Homozygous animals exhibited reduced primitive erythropoiesis and died around embryonic day 13, owing to failure of definitive fetal liver erythropoiesis. Both types of mutations exhibited identical phenotypes, indicating that EPO and the EPOR are crucial for definitive erythropoiesis in vivo and that no other ligands or receptors can replace them. Committed erythroid BFU-E and CFU-E progenitors were present in both homozygous fetal livers. Thus, neither EPO nor the EPOR is required for erythroid lineage commitment or for the proliferation and differentiation of BFU-E to CFU-E progenitors. EPO and the EPOR are crucial in vivo for the proliferation and survival of CFU-E progenitors and their irreversible terminal differentiation.

Animals↗
PubMed1995

Loss of methylation activates Xist in somatic but not in embryonic cells.

The mouse Xist gene, which is expressed only from the inactive X chromosome, is thought to play a role in the initiation of X inactivation. The 5' end of this gene is fully methylated on the active X chromosome and completely demethylated on the inactive X chromosome, suggesting that DNA methylation may be involved in controlling allele-specific transcription of this gene. To directly investigate the importance of DNA methylation in the control of Xist expression, we have examined its methylation patterns and expression in ES cells and embryos that are deficient in DNA methyltransferase activity. We report here that demethylation of the Xist locus in male mutant embryos induces Xist expression, thus establishing a direct link between demethylation and expression of the Xist gene in the postgastrulation embryo. The transcriptional activity of Xist in undifferentiated ES cells, however, appears to be independent of its methylation status. These results suggest that methylation may only become essential for Xist repression after ES cells have differentiated or after the embryo has undergone gastrulation.

Animals↗
PubMed1995

Sensory but not motor neuron deficits in mice lacking NT4 and BDNF.

Neurotrophins play important roles in neuronal survival during vertebrate development. Neurotrophin-4 (NT4), alone or in combination with brain-derived neurotrophic factor (BDNF), has been suggested to be necessary for the survival of peripheral sensory neurons and central nervous system (CNS) neurons, including motor neurons. To define the role of NT4 in vivo, we generated mice lacking NT4 by gene targeting. NT4-deficient mice were viable but exhibited a loss of sensory neurons in the nodose-petrosal and geniculate ganglia. In contrast, motor neurons of the facial nucleus and sympathetic neurons of the superior cervical ganglion were unaffected, and there was no obvious loss of dopaminergic neurons in the substantia nigra. In mice lacking both NT4 and BDNF, facial neurons remained unaffected, whereas the loss of sensory neurons was more severe than with either mutation alone. Thus NT4 is required during development for the survival of some peripheral sensory neurons but not sympathetic or motor neurons.

Animals↗
PubMed1995

Suppression of intestinal neoplasia by DNA hypomethylation.

We have used a combination of genetics and pharmacology to assess the effects of reduced DNA methyltransferase activity on ApcMin-induced intestinal neoplasia in mice. A reduction in the DNA methyltransferase activity in Min mice due to heterozygosity of the DNA methyltransferase gene, in conjunction with a weekly dose of the DNA methyltransferase inhibitor 5-aza-deoxycytidine, reduced the average number of intestinal adenomas from 113 in the control mice to only 2 polyps in the treated heterozygotes. Hence, DNA methyltransferase activity contributes substantially to tumor development in this mouse model of intestinal neoplasia. Our results argue against an oncogenic effect of DNA hypomethylation. Moreover, they are consistent with a role for DNA methyltransferase in the generation of the C to T transitions seen at high frequency in human colorectal tumors.

Adenomatous Polyposis Coli↗
PubMed1995

Functional analysis of activins during mammalian development.

Activins are dimeric (beta A beta A; beta B beta B; beta A beta B) members of the transforming growth factor-beta superfamily. They are widely expressed during murine development, are highly conserved during vertebrate evolution, and may be involved in mesoderm induction and neurulation in Xenopus laevis and Oryzias latipes. To investigate the function of mammalian activins in vivo, we generated mice with mutations either in activin-beta A or in both activin-beta A and activin-beta B. Activin-beta A-deficient mice develop to term but die within 24 h of birth. They lack whiskers and lower incisors and have defects in their secondary palates, including cleft palate, demonstrating that activin-beta A must have a role during craniofacial development. Mice lacking both activin subunits show the defects of both individual mutants but no additional defects, indicating that there is no functional redundancy between these proteins during embryogenesis. In contrast to observations in lower vertebrates, zygotic expression of activins is not essential for mesoderm formation in mice.

Activins↗
PubMed1995

The MyoD family of transcription factors and skeletal myogenesis.

Gene targeting has allowed the dissection of complex biological processes at the genetic level. Our understanding of the nuances of skeletal muscle development has been greatly increased by the analysis of mice carrying targeted null mutations in the Myf-5, MyoD and myogenin genes, encoding members of the myogenic regulatory factor (MRF) family. These experiments have elucidated the hierarchical relationships existing between the MRFs, and established that functional redundancy is a feature of the MRF regulatory network. Either MyoD or Myf-5 is sufficient for the formation or survival of skeletal myoblasts. Myogenin acts later in development and plays an essential in vivo role in the terminal differentiation of myotubes.

Animals↗
PubMed1995

Suppressed epileptogenesis in BDNF mutant mice.

Kindling is an animal model of epilepsy in which repeated electrical stimulations lead to progressive and permanent amplification of seizure activity, culminating in generalized convulsions. Each brief period of seizure activity during kindling epileptogenesis causes a marked, transient increase of the synthesis of brain-derived neurotrophic factor (BDNF) in cortical and hippocampal neurons. We find that the development of kindling is markedly suppressed in mice heterozygous for a deletion of the BDNF gene. In contrast, the maintenance of kindling is unaffected. The mutant mice show lower levels of BDNF mRNA in cortical and hippocampal neurons after seizures than do wild-type mice. Hippocampal mossy fiber sprouting is augmented in BDNF mutants but there are no other morphological abnormalities. These results show that BDNF plays an important role in establishing hyperexcitability during epileptogenesis, probably by increasing efficacy in stimulated synapses.

Animals↗
PubMed1995

Complementary roles of BDNF and NT-3 in vestibular and auditory development.

The physiological role of BDNF and NT-3 in the development of the vestibular and auditory systems was investigated in mice that carry a deleted BDNF and/or NT-3 gene. BDNF was the major survival factor for vestibular ganglion neurons, and NT-3, for spiral ganglion neurons. Lack of BDNF and NT-3 did not affect ingrowth of nerve fibers into the vestibular epithelium, but BDNF mutants failed to maintain afferent and efferent innervation. In the cochlea, BDNF mutants lost type 2 spiral neurons, causing an absence of outer hair cell innervation. NT-3 mutants showed a paucity of afferents and lost 87% of spiral neurons, presumably corresponding to type 1 neurons, which innervate inner hair cells. Double mutants had an additive loss, lacking all vestibular and spiral neurons. These results show that BDNF and NT-3 are crucial for inner ear development and, although largely coexpressed, have distinct and nonoverlapping roles in the vestibular and auditory systems.

Afferent Pathways↗
PubMed1995

Differential role of the low affinity neurotrophin receptor (p75) in retrograde axonal transport of the neurotrophins.

The receptor mechanisms mediating the retrograde axonal transport of the neurotrophins have been investigated in adult rats. We show that transport of the TrkB ligands NT-4 and BDNF to peripheral neurons is dependent on the low affinity neurotrophin receptor (LNR). Pharmacological manipulation of LNR in vivo using either an anti-LNR antibody or a soluble recombinant LNR extracellular domain completely blocked retrograde transport of NT-4 and BDNF to sensory neurons, while having minimal effects on the transport of NGF in either sensory or sympathetic neurons. Furthermore, in mice with a null mutation of LNR, the transport of NT-4 and BDNF, but not NGF, was dramatically reduced. These observations demonstrate a selective role for LNR in retrograde transport of the various neurotrophins from distinct target regions in vivo.

Animals↗
PubMed1995

Targeted mutation in the col5a2 gene reveals a regulatory role for type V collagen during matrix assembly.

The tissue-specific organization of collagen molecules into tridimensional macroaggregates determines the physiomechanical properties of most connective tissues, but the factors and mechanisms controlling this process are unknown. It has been postulated that quantitatively minor types V and XI collagen regulate the growth of type I and II collagen fibrils, respectively. To test this hypothesis, we created mice that produce a structurally abnormal alpha 2(V) collagen chain. Homozygous mutant mice survive poorly, possibly because of complications from spinal deformities, and exhibit skin and eye abnormalities caused by disorganized type I collagen fibrils. Our results demonstrate that type V collagen is a key determinant in the assembly of tissue-specific matrices, and provide an animal model for human connective tissue disorders.

Animals↗
PubMed1995

A targeted mutation at the known collagenase cleavage site in mouse type I collagen impairs tissue remodeling.

Degradation of type I collagen, the most abundant collagen, is initiated by collagenase cleavage at a highly conserved site between Gly775 and Ile776 of the alpha 1 (I) chain. Mutations at or around this site render type I collagen resistant to collagenase digestion in vitro. We show here that mice carrying a collagenase-resistant mutant Col1a-1 transgene die late in embryo-genesis, ascribable to overexpression of the transgene, since the same mutation introduced into the endogenous Col1a-1 gene by gene targeting permitted normal development of mutant mice to young adulthood. With increasing age, animals carrying the targeted mutation developed marked fibrosis of the dermis similar to that in human scleroderma. Postpartum involution of the uterus in the mutant mice was also impaired, with persistence of collagenous nodules in the uterine wall. Although type I collagen from the homozygous mutant mice was resistant to cleavage by human or rat fibroblast collagenases at the helical site, only the rat collagenase cleaved collagen trimers at an additional, novel site in the nonhelical N-telopeptide domain. Our results suggest that cleavage by murine collagenase at the N-telopeptide site could account for resorption of type I collagen during embryonic and early adult life. During intense collagen resorption, however, such as in the immediate postpartum uterus and in the dermis later in life, cleavage at the helical site is essential for normal collagen turnover. Thus, type I collagen is degraded by at least two differentially controlled mechanisms involving collagenases with distinct, but overlapping, substrate specificities.

Amino Acid Sequence↗
PubMed1995

Studies on the physiological role of brain-derived neurotrophic factor and neurotrophin-3 in knockout mice.

Brain-derived neurotrophic factor and neurotrophin-3 deficient mice were generated by gene targeting. The analysis of these mice has led to the characterization of their role in the survival of neurons in the peripheral nervous system. NT-3 deficient mice displayed severe movement defects and most died shortly after birth. The mutation causes loss of substantial portions of cranial and spinal peripheral sensory and sympathetic neurons. Significantly, spinal proprioceptive afferents and their peripheral sense organs (muscle spindles and Golgi tendon organs) were completely absent in homozygous mutant mice. BDNF deficient mice displayed deficiencies in coordination and balance. Excessive loss of neurons was detected in most of the peripheral sensory ganglia examined, but the survival of sympathetic neurons was not affected. The most marked reduction of neurons was observed in the vestibular ganglion, leading to a loss of innervation of the sensory epithelia of the vestibular compartments of the inner ear.

Afferent Pathways↗
PubMed1994

Toxicity of 5-aza-2'-deoxycytidine to mammalian cells is mediated primarily by covalent trapping of DNA methyltransferase rather than DNA demethylation.

The deoxycytidine analog 5-aza-2'-deoxycytidine (5-azadCyd) has been widely used as a DNA methylation inhibitor to experimentally induce gene expression and cellular differentiation. Prior to the availability of mutant mice with altered DNA methyltransferase levels, treatment of cells with drugs has been the only means to experimentally manipulate the level of genomic DNA methylation in mammalian cells. Substitution of DNA with 5-azadCyd leads to covalent trapping of the enzyme, thereby depleting the cells of enzyme activity and resulting in DNA demethylation. 5-AzadCyd or 5-azacytidine treatment causes multiple changes in treated cells, including activation of silent genes, decondensation of chromatin, and induction of cellular differentiation, all of which are believed to be consequences of drug-induced demethylation. 5-AzadCyd is highly toxic in cultured cells and animals and is utilized as a potent antitumor agent for treatment of certain human cancers. It has been postulated that the toxicity of the drug in mammalian cells is also due to its inhibition of DNA methylation. The chemistry of the methylation reaction is consistent, however, with an alternative mechanism: the cytotoxic effect of 5-azadCyd may be directly mediated through the covalent binding of DNA methyltransferase to 5-azadCyd-substituted DNA. We have tested this possibility by using embryonic stem cells and mice with reduced levels of DNA methyltransferase due to a targeted mutation of the gene. When exposed to 5-azadCyd mutant embryonic stem cells or embryos were significantly more resistant to the toxic effects of the drug than wild-type cells and embryos, respectively. These results strongly suggest that the cellular DNA methyltransferase itself, rather than the secondary demethylation of genomic DNA, is the primary mediator of 5-azadCyd cytotoxicity. In light of our results, some conclusions from previous studies using 5-azadCyd in order to experimentally manipulate cellular methylation levels may have to be reassessed. Also, our data make clear predictions for cancer treatment: tumor cells with elevated DNA methyltransferase levels would be expected to be susceptible to treatment with 5-azadCyd, whereas tumors with reduced levels of the enzyme would be resistant.

Animals↗
PubMed1994

Mice lacking alpha 1 (IX) collagen develop noninflammatory degenerative joint disease.

Type IX collagen is a nonfibrillar collagen composed of three gene products, alpha 1(IX), alpha 2(IX), and alpha 3(IX). Type IX molecules are localized on the surface of type II-containing fibrils and consist of two arms, a long arm that is crosslinked to type II collagen and a short arm that projects into the perifibrillar space. In hyaline cartilage, the alpha 1(IX) collagen transcript encodes a polypeptide with a large N-terminal globular domain (NC4), whereas in many other tissues an alternative transcript encodes an alpha 1(IX) chain with a truncated NC4 domain. It has been proposed that type IX molecules are involved in the interaction of fibrils with each other or with other components of the extracellular matrix. To test this hypothesis, we have generated a mouse strain lacking both isoforms of the alpha 1(IX) chain. Homozygous mutant mice are viable and show no detectable abnormalities at birth but develop a severe degenerative joint disease resembling human osteoarthritis.

Animals↗
PubMed1994

Lack of neurotrophin-3 leads to deficiencies in the peripheral nervous system and loss of limb proprioceptive afferents.

Neurotrophin-3-deficient (NT-3-deficient) mice were generated by gene targeting. Mutant mice displayed severe movement defects of the limbs, and most died shortly after birth. Substantial portions of peripheral sensory and sympathetic neurons were lost while motor neurons were not affected. Significantly, spinal proprioceptive afferents and their peripheral sense organs (muscle spindles and Golgi tendon organs) were completely absent in homozygous mutant mice. This correlated with a loss of parvalbumin and carbonic anhydrase-positive neurons in the dorsal root ganglion. No gross abnormalities were seen in Pacinian corpuscles, cutaneous afferents containing substance P and calcitonin gene-related peptide, and deep nerve fibers in the joint capsule and tendon. Importantly, the number of muscle spindles in heterozygous mutant mice was half of that in control mice, indicating that NT-3 is present at limiting concentrations in the embryo.

Animals↗