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D W Pfaff

Publications and source records attributed to D W Pfaff.

At least 127 records · Page 7Linked to original sources
PubMed1994

In vitro electrophysiological characterization of midbrain periaqueductal gray neurons in female rats: responses to GABA- and Met-enkephalin-related agents.

Met-Enkephalin, which can be induced by estrogen in the ventromedial nucleus of hypothalamus (VMH), has been proposed to help mediate estrogenic action on lordosis behavior by acting on midbrain periaqueductal gray (PAG) neurons. Also, in the PAG, GABA may locally regulate the levels of lordosis behavior through GABAA receptors. Therefore, we examined the effects of both Met-enkephalin and GABA-related agents on neuronal activity of PAG neurons in slices. Overall, 72.6% of the PAG neurons were inhibited by GABA and 60.9% of GABA-responsive neurons were also excited by the GABAA receptor antagonist, bicuculline methiodide (BMI), suggesting that many of GABA-responsive PAG neurons are tonically inhibited by GABAergic neurons through GABAA receptors. Dorsal PAG neurons were more responsive to BMI than ventral PAG neurons. Moreover, in the middle part of the dorsal PAG, where prominent inhibitory behavioral effects of BMI have been reported, BMI excited 94% of GABA-responsive PAG neurons from estrogen-treated animals, significantly more than observed in ovariectomized control (50%). The most frequent action of Met-enkephalin on PAG neurons was inhibitory (38 out of 149 recorded neurons) although it excited 12 neurons. A dose-dependent increase of inhibitory action of enkephalin was found in the estrogen-primed group but not in the ovariectomized control group while higher doses of enkephalin failed to excite any more neurons in both groups. Most frequently (90%), enkephalin inhibited the same neurons as those on which GABA had the inhibitory effects. Conversely, these neurons composed about 50% of the entire GABA-responsive PAG neurons. Moreover, 76% of neurons inhibited by enkephalin were found to be tonically inhibited by endogenous GABA through GABAA receptors. It is argued, therefore, that increased enkephalinergic influences from the VMH to the PAG in estrogen-treated females could participate in the PAG neuronal control of lordosis by acting on the same neurons as are innervated by intrinsic GABAergic neurons. Since GABAA agonists actually facilitate lordosis in the PAG, these PAG neurons inhibited by both GABA and enkephalin may themselves facilitate behaviors which are antagonistic to lordosis, such as defensive behaviors.

Animals↗
PubMed1994

Activation of protein kinase C in the hypothalamic ventromedial nucleus or the midbrain central gray facilitates lordosis.

Many neurotransmitters and neuropeptides can act through the hypothalamic ventromedial nucleus (VMN) or midbrain central gray (MCG) to facilitate lordosis. Since these lordosis-facilitating agents can also stimulate the phosphoinositide (PI) second-messenger pathway, it was hypothesized that direct activation of this pathway can also potentiate the behavior. To evaluate this possibility, a phorbol ester, TPA (12-O-tetradecanoyl phorbol 13-acetate), was used to activate a key enzyme, protein kinase C (PKC), of the PI pathway in ovariectomized (OVX) rats either primed or not primed with estrogen. These female rats were paired with males for mating tests before and after an intracerebral infusion of TPA, and both the lordosis quotient (LQ) and the lordosis strength (LS) were measured. Bilateral infusion of TPA (5 micrograms/0.5 microliter or 0.2 microgram/0.2 microliter, but not 0.1 microgram/0.2 microliter/side) into the VMN or MCG of estrogen-primed subjects facilitated both LQ and LS in 30 min, peaked at 60-90 min, and the facilitation lasted for more than 180 min. This facilitatory effect of TPA was: (1) not observed in OVX rats not primed with estrogen; (2) not observed if the infused TPA did not reach both sides of the VMN or MCG; (3) not mimicked by 4 alpha-phorbol 12,13-didecanoate, which does not activate PKC; (4) blocked by PKC inhibitors (H7 10 mM or staurosporine 1 microM, 0.2 microliter/side), which by themselves did not facilitate lordosis; and (5) was not affected by pretreatment of the progestin antagonist RU486.(ABSTRACT TRUNCATED AT 250 WORDS)

Alkaloids↗
PubMed1994

Preproenkephalin promoter yields region-specific and long-term expression in adult brain after direct in vivo gene transfer via a defective herpes simplex viral vector.

We have previously used a defective herpes simplex virus vector to express a foreign gene in the adult rat brain. One application of this technology would be the in vivo analysis of promoter function in brain after de novo transfer, which would allow the rapid generation of vectors with localized application in a broad range of mammalian species while avoiding influences of other nearby promoters. A 2.7-kb fragment of the rat preproenkephalin promoter was placed upstream of the bacterial lacZ gene in our herpes simplex virus amplicon. A restricted pattern of lacZ expression was observed in vivo, which follows previously observed patterns of endogenous preproenkephalin expression. These results, from the direct gene transfer into an adult animal brain for in vivo promoter analysis, demonstrate that sequence information that influences restricted expression of preproenkephalin is located within 2.7 kb upstream of transcriptional initiation. lacZ expression was also observed in rat brain for 2 months after direct transfer, and PCR analysis confirmed the continued presence of amplicon DNA in lacZ-positive sections. Restricted and long-term expression observed with an endogenous promoter has important implications for gene therapy using viral vectors.

Animals↗
PubMed1994

Antibody to neural cell adhesion molecule can disrupt the migration of luteinizing hormone-releasing hormone neurons into the mouse brain.

The neurons which synthesize and release luteinizing hormone-releasing hormone (LHRH), are hypothesized to originate in the epithelium of the medial olfactory pit and to migrate into the brain along a scaffolding made up of neural cell adhesion molecule (NCAM)-immunoreactive branches of the terminal and vomeronasal nerves. These LHRH neurons, studied by immunocytochemical and autoradiographic procedures, were found to originate within a very short period of embryogenesis, specifically day 10, in mice, and to follow a remarkably ordered spatiotemporal course along the migration route into the brain. The purpose of the present experiments was to determine whether perturbation of the NCAM-immunoreactive migration route, at a particular time in development, would arrest the migration of LHRH neurons into the brain. We found that a 1 microliter injection of antiserum to NCAM into the area of the olfactory pit, on day 10 of embryogenesis, significantly reduced the number of LHRH-immunoreactive neurons seen in the epithelium of the medial olfactory pit, with a concomitant significant reduction in the number of LHRH-immunoreactive cells seen outside of the placode, on the migration route. These results confirm our initial hypothesis that LHRH neurons migrate from the epithelium of the olfactory pit to the brain and indicate that NCAM plays a causal role in this phenomenon.

Animals↗
PubMed1994

In situ hybridization detection of trkA mRNA in brain: distribution, colocalization with p75NGFR and up-regulation by nerve growth factor.

In situ hybridization techniques were used to examine the distribution and the nerve growth factor (NGF) regulation of trkA mRNA in the adult rat brain in order to identify neurons in discrete regions of the brain that may be NGF responsive. In agreement with previous studies, trkA mRNA was detected within cells located in the medial septum (MS), diagonal band of Broca (DBB), and caudate. trkA mRNA was also detected in many other regions of the brain, including the nucleus basalis of Meynert, substantia innominata, paraventricular nucleus of the thalamus, interpeduncular nucleus, prepositus hypoglossal nucleus, vestibular nuclei, raphe obscuris, cochlear nucleus, sensory trigeminal nuclei, and gigantocellular as well as perigigantocellular neurons in the medullary reticular formation. By combining in situ hybridization detection of trkA mRNA with immunocytochemical detection of p75NGFR, it was determined that the vast majority (> 90%) of the trkA mRNA-containing cells detected in the MS and DBB also express p75NGFR. Likewise, the vast majority of p75NGFR-IR cells detected in the MS and DBB expressed trkA mRNA. Intracerebroventricular infusions of NGF into the third ventricle adjacent to the preoptic area resulted in a 58% increase in relative cellular levels of trkA mRNA in the horizontal limb of the DBB. These data provide evidence that both p75NGFR and trkA are expressed by NGF-responsive neurons in the MS and DBB. In addition, we note that areas that contained trkA mRNA and that also have been reported to contain p75NGFR are areas where high-affinity NGF binding sites have been observed autoradiographically, whereas areas that contain either trkA or p75NGFR alone are areas where no high-affinity NGF binding has been reported. Together, these findings suggest that both trkA and p75NGFR play an important role in the formation of high-affinity NGF receptors in brain and, furthermore, suggest that NGF may have physiological effects within many regions of the brain outside of the basal forebrain.

Animals↗
PubMed1994

Intracerebral administration of antisense oligodeoxynucleotides to GAD65 and GAD67 mRNAs modulate reproductive behavior in the female rat.

Increased GABA activity in the medial hypothalamus (HYP) and midbrain central gray (MCG), but not the preoptic area (POA), facilitates sexual receptivity in the female rat [40]. In the current experiments, ovariectomized females were chronically treated with estrogen (via silastic capsules) to maintain a continuously high level of lordosis response. Administration of crystalline antisense oligodeoxynucleotide to the GABA synthetic enzyme, GAD67, into the HYP and MCG significantly and reversibly reduced lordosis response for 1-2 days, but did not inhibit lordosis when administered into the POA. Administration of a control oligonucleotide, consisting of the same nucleotide bases but in a scrambled sequence, did not significantly modulate behavior when infused into any brain areas. When oligodeoxynucleotide antisense to GAD67 was suspended in oil and then infused into the HYP or MCG it was more effective and resulted in less inter-animal variability. Subsequent experiments involving infusions into the MCG compared the effectiveness of antisense oligonucleotides to the two different forms of GAD, known as GAD65 and GAD67. Oligodeoxynucleotides antisense to the mRNA for either gene were effective at reducing lordosis behavior but with a different time course. Oligonucleotide antisense to GAD67 significantly reduced behavior within 24 h of infusion and there was full recovery by 4 days post-infusion. GAD65 antisense oligonucleotide did not significantly reduce behavior until 48 h post infusion and animals did not fully recover to pretest levels of lordosis until 5 days post-infusion. When antisense oligonucleotide for the two genes was administered simultaneously, the inhibition of lordosis was maximal at 24 h and stayed depressed for 4 days. There did not appear to be an additive effect of the two different antisense oligonucleotides when administered together. Tissue GABA levels in HYP and MCG of individual rats assayed by HPLC were no longer correlated with lordosis score after antisense oligonucleotide infusion but were after infusions of scrambled control oligos. Immunoblotting for the two forms of GAD revealed that GAD67 antisense oligonucleotide infusion led to significant decreases in both GAD67 and GAD65 protein levels as compared to infusions of scrambled control oligo. In addition, the levels of a neuronal marker, neuron-specific enolase, also decreased (although nonsignificantly) suggesting either a temporary shutdown of protein synthesis or a degeneration of GABAergic neurons after GAD67 antisense oligonucleotide infusion.

Animals↗
PubMed1994

Effects of estrogen replacement on the relative levels of choline acetyltransferase, trkA, and nerve growth factor messenger RNAs in the basal forebrain and hippocampal formation of adult rats.

Previous studies have shown that estrogen replacement can significantly affect the expression of choline acetyltransferase immunoreactivity (ChAT-IR) and low-affinity (p75NGFR) nerve growth factor receptors within cholinergic neurons located in specific regions of the basal forebrain. To extend this work, we have examined the effects of estrogen replacement on relative levels of choline acetyltransferase (ChAT), trkA, and nerve growth factor (NGF) mRNAs within different regions of the basal forebrain and hippocampal formation using quantitative in situ hybridization techniques. Ovariectomized Sprague-Dawley rats received continuous estrogen replacement for 2 days, 1 week, or 2 weeks. The data show that estrogen replacement results in significant increases in relative cellular levels of ChAT mRNA in the medial septum (MS) and nucleus basalis magnocellularis (nBM), but not in the horizontal limb of the diagonal band of Broca (HDB) or the striatum. In contrast, estrogen replacement resulted in significant decreases in relative levels of NGF mRNA in the hippocampus and of trkA mRNA in the MS and nBM (but not in the HDB or the striatum). The time-course of these effects is consistent with estrogen having a direct effect on ChAT expression which is followed by indirect effects on trkA. The data are also consistent with previous findings in which estrogen replacement resulted in significant increases in ChAT-IR which were followed by significant decreases in p75NGFR mRNA and protein and then a reduction in ChAT-IR back to control levels. Together, these data indicate that estrogen replacement can have significant effects on basal forebrain cholinergic function, and that some of these effects may be mediated by effects of estrogen replacement on the expression of NGF and NGF receptors.

Animals↗
PubMed1994

Specific patterns of defective HSV-1 gene transfer in the adult central nervous system: implications for gene targeting.

Viral vectors are a means by which genes can be delivered to specific sites in the adult central nervous system. Nevertheless, the interaction between the viral vector and cells of the nervous system, which forms the basis for specific gene transfer, is not well understood. In this study a nonreplicating defective herpes simplex virus type 1 vector, expressing the marker gene lacZ, was stereotaxically injected at varying titers into the rat central nervous system. Three sites were targeted: the caudate nucleus, dentate gyrus, and cerebellar cortex, and the resulting patterns of beta-galactosidase activity were examined. Many cells of neuronal and glial morphology, and of differing neuronal subtypes, expressed beta-galactosidase at each of the injection sites. However, beta-galactosidase activity was also detected in distant secondary brain areas, the neurons of which make afferent connections with the primary sites. This strongly suggested that the retrograde transport of defective virus was the basis for the enzyme activity observed at a distance. Moreover, retrograde transport to secondary sites was found to be highly selective and restricted to certain retrograde neuroanatomical pathways in a specific and titer dependent fashion. The pathways observed were predominantly, but not exclusively, monoaminergic in origin. This finding is supported by reports of specific tropism by HSV for monoaminergic circuits in experimental encephalitis and transneuronal tracing studies. Our observations suggest that certain functional neuronal populations, which are permissive for the retrograde transfer of defective HSV-1 vectors, might be specifically targeted for gene transfer using this approach. Conversely, a knowledge of the pathways permissive for viral uptake, retrograde transfer, and subsequent gene expression will be essential in order to predict the consequences of gene transfer using viral vectors.

Animals↗
PubMed1994

Mutant herpes simplex virus induced regression of tumors growing in immunocompetent rats.

Herpes simplex virus (HSV) mutants kill dividing tumor cells but spare non-proliferating, healthy brain tissue and may be useful in developing new treatment strategies for malignant brain tumors. Two HSV mutants, a thymidine kinase deficient virus (TK-) and a ribonucleotide reductase mutant (RR-), killed 7/7 human tumor cell lines in tissue culture. The TK-HSV killed Rat RG2 glioma and W256 carcinoma lines but not the rat C6 glioma in culture. TK-HSV replication (12 pfu/cell) was similar to wild-type HSV (10 pfu/cell) in rapidly dividing W256 cells in tissue culture, but was minimal (< 1 pfu/cell) in serum-starved cells, suggesting that the proliferative activity of tumor cells at the site and time of TK-HSV injection may influence efficacy in vivo. Subcutaneous W256 tumors in male Sprague-Dawley rats were injected with TK-HSV or free inoculum. A significant effect of TK-HSV therapy on W256 tumor growth was demonstrated compared to controls (p = 0.002). Complete regression was observed in 4/9 experimental tumors, with no recurrence over 6 months. Tumor growth in the remaining 5/9 animals was attenuated during the first 3 to 5 days after treatment, but not beyond 5 days compared to 9 matched control animals; no tumor regression was observed in any of the control animals. These results suggest that HSV mutants are potentially useful as novel therapeutic agents in the treatment of tumors in immunocompetent subjects.

Animals↗
PubMed1994

Roles of second-messenger systems and neuronal activity in the regulation of lordosis by neurotransmitters, neuropeptides, and estrogen: a review.

Many neurotransmitters and neuropeptides can affect the rodent feminine sexual behavior, lordosis, when administered in the ventromedial hypothalamus (VMH), midbrain central gray (MCG), or other brain regions. A survey of the electrophysiological and biochemical actions of these neural agents revealed that there is a very consistent association between lordosis facilitation with both the activation of the phosphoinositide (PI) pathway and the excitation of VMH and MCG neurons. In contrast, lordosis inhibition is associated, less consistently, with alterations of the adenylate cyclase (AC) system and the inhibition of neuronal activity. The findings that lordosis could be facilitated by going beyond membrane receptors and directly activating the PI pathway, suggest that this second-messenger pathway is a common mediator for the lordosis-facilitating agents. Furthermore, as in the case of stimulating membrane receptors, direct activation of this common mediator also requires estrogen priming for lordosis facilitation. Therefore, it is likely that the PI pathway is modulated by estrogen in the permissive action of estrogen priming. Indeed, a literature review shows that estrogen can affect selective isozymes of key enzyme families of the PI pathway at various levels. Such selective modulations, at several levels, could easily alter the course of a PI cascade; thence, the eventual functional outcome. These findings prompt us to propose that estrogen enables lordosis to be facilitated by a selective modulation of the PI pathway.

Animals↗
PubMed1994

Expression of the growth-associated protein B-50/GAP43 via a defective herpes-simplex virus vector results in profound morphological changes in non-neuronal cells.

This study describes the creation and application of a defective herpes simplex viral (HSV) vector for B-50/GAP-43, a neural growth-associated phosphoprotein. We demonstrate abundant expression of B-50/GAP-43 in cultured non-neuronal cells (African green monkey kidney cells [vero cells] and Rabbit skin cells) via this HSV vector. When B-50/GAP-43 was expressed in non-neuronal cells major morphological changes occurred that included extensive membrane ruffling, the formation of filopodia and long thin extensions reminiscent of neurites. These extensions often terminated in growth cone-like structures. Quantitation of these morphological changes at different times following infection demonstrates that the surface area of the B-50/GAP-43-expressing cells started to increase between 6 and 10 h post-infection. At 72 h, B-50/GAP-43-positive cells were 3.0 times larger in size and one third of the cells expressed long processes with a mean length of 165 +/- 14.5 microns. Ultrastructural studies of cells 48 h after infection revealed that B-50/GAP-43 is predominantly localized at the plasma membrane of the elaborated processes. Some immunoreactivity was associated with vesicular structures that appear to be in-transit in the processes. These observations suggest that B-50/GAP-43 acts at the plasmamembrane to induce a neuron-like morphology in non-neuronal cells persisting for several days in culture. In the future the defective viral vector will enable gene transfer to express B-50/GAP-43 in neurons in vivo in order to study its involvement in regenerative sprouting and neuroplasticity.

Animals↗
PubMed1994

Linear dimeric interleukin-2 obtained by the use of a defective herpes simplex viral vector: conformation-activity relationship.

An interleukin-2 dimer, produced enzymatically by a nerve-derived transglutaminase in vitro, is cytotoxic to oligodendrocytes, unlike the immune-derived monomeric interleukin-2. The object of this study was to establish a way to produce a dimer of interleukin-2 in quantities, by means of genetic engineering, and to confirm that the structure of the resulting molecule is critical for its function. A defective herpes simplex virus vector was utilized for overproduction of a dimeric interleukin-2. The resulting linear dimer, which is a translational product, differs from the enzymatically produced dimer, which is a posttranslational modification of interleukin-2. The linear dimer, while retaining the known interleukin-2 activity of monomeric interleukin-2 with respect to mitogenicity on T cells, was not cytotoxic to oligodendrocytes. This finding suggests that the lack of cytotoxicity of the linear dimeric interleukin-2 is not caused by a loss of activity during its preparation but is related to its conformational structure, which evidently does not meet the requirements for cytotoxicity. This study opens the way to the design at the transcriptional level of modified proteins and their efficient production, provided that the new transcript encodes for the desired modification in the protein at the appropriate sites.

Animals↗
PubMed1994

Activation of the transcription factor NF-KB in GH3 pituitary cells.

Since several genes expressed in the pituitary can bind the transcription factor NF-KB, its presence and regulation was examined in the GH3 pituitary cell line. An electrophoretic mobility shift assay using nuclear extracts and an oligonucleotide probe corresponding to the Ig KB binding site was employed to identify activated NF-KB. One complex possessed properties characteristic of NF-KB: co-migration with an NF-KB complex and binding specificity restricted to NF-KB binding DNA sequences. Antibodies to the NF-KB subunits NFKB1p50 (p50) and RelA (p65) interacted with the extract-DNA complex. Activation of NF-KB in GH3 cells was increased by PMA or the cytokine tumor necrosis factor alpha. A synergy between PMA and TNF or a calcium mobilizing agent was seen in NF-KB activation. Further TNF activation was enhanced by TRH. These observations indicate the presence of NF-KB in GH3 cells and demonstrate its activation by hormones/second messengers that act on pituitary cells.

Base Sequence↗
PubMed1994

Competition for DNA steroid response elements as a possible mechanism for neuroendocrine integration.

For the analysis of a simple steroid-dependent mating behavior, careful response definition, complete neural circuit delineation and placement of estrogen-responsive cells within this circuit have been accomplished. Molecular studies of two relevant genes have emphasized DNA/RNA hybridization assays and DNA binding techniques. For both the rat preproenkephalin gene and the gene for the progesterone receptor, a strong induction by estrogen, tissue specificity of expression and a sex difference in regulation are prominent phenomena. On the rat preproenkephalin promoter, estrogen (ER) and thyroid receptors may compete for a DNA binding site. Likewise, progesterone (PR) and glucocorticoid receptors may compete for the same sites. On the rat PR gene, interactions between ER and AP-1 binding proteins are of special interest. Such interactions could underlay competitions and synergies between steroid hormones and neurally signalled events in the environment.

Animals↗
PubMed1994

Long-term gene expression and phenotypic correction using adeno-associated virus vectors in the mammalian brain.

Adeno-associated viral (AAV) vectors are non-pathogenic, integrating DNA vectors in which all viral genes are removed and helper virus is completely eliminated. To evaluate this system in the post-mitotic cells of the brain, we found that an AAV vector containing the lacZ gene (AAVlac) resulted in expression of beta-galactosidase up to three months post-injection in vivo. A second vector expressing human tyrosine hydroxylase (AAVth) was injected into the denervated striatum of unilateral 6-hydroxydopamine-lesioned rats. Tyrosine hydroxylase (TH) immunoreactivity was detectable in striatal neurons and glia for up to four months and we also found significant behavioural recovery in lesioned rats treated with AAVth versus AAVlac controls. Safe and stable TH gene transfer into the denervated striatum may have potential for the genetic therapy of Parkinson's disease.

Adenoviruses, Human↗
PubMed1994

Disruption of estrous cyclicity following administration of a luteinizing hormone-releasing hormone antagonist to the preoptic area of the rat.

Experiments were designed to test the hypothesis that LHRH receptors in the preoptic area (POA) are of physiological importance for maintaining estrous cyclicity in the rat. Bilateral cannulae were implanted just dorsal to the POA. Estrous cycles were monitored daily by vaginal smears. Antide, a long-acting LHRH antagonist, was infused bilaterally (2.5 micrograms/side) in the POA or the hypothalamus on the mornings of diestrus I and II. As controls, at separate times, rats also received similar infusions of either vehicle (1:1 water:propylene glycol) or a bombesin antagonist (#B0650; Sigma, St. Louis, MO). Collection of daily vaginal smears continued, and the number of days from the first infusion to the next day of estrus that preceded a normal cycle was recorded. After infusion of Antide into the POA, rats demonstrated varying durations of interrupted cycles ranging from 11 days to more than 100 days. These periods of disruption were characterized by either long periods of diestrus, long periods of estrus, or an extended period of diestrus followed by an extended period of estrus. After infusion of Antide into the dorsomedial, ventromedial, or anterior hypothalamic areas, rats had either a 4- or 5-day estrous cycle and continued to cycle normally. Likewise, infusions into the septum had no effect. Infusion of vehicle or bombesin antagonist into any of the hypothalamic or POA sites tested also resulted in no interruptions in the cyclic activity of the rats. Therefore, it appears that functional LHRH receptors in the POA are necessary to drive the normal estrous cycle.

Animals↗
PubMed1994

Luteinizing hormone-releasing hormone receptor messenger ribonucleic acid expression in the rat pituitary during lactation and the estrous cycle.

To study mechanisms underlying the modulation of luteinizing hormone-releasing hormone receptor (LHRH-R) during lactation and the estrous cycle, we used a reverse transcriptase-polymerase chain reaction (RT-PCR) procedure to generate a probe for rat LHRH-R messenger RNA (mRNA). Using primers based on the mouse sequence, we amplified an approximately 300 bp fragment from rat pituitary complementary DNA. This PCR product was shown to be part of LHRH-R cDNA by direct sequencing and by comparing to the rat LHRH-R cDNA reported recently. Then, this PCR fragment was used as a probe for northern blotting analysis. The level of LHRH-R mRNA in the pituitary was significantly decreased during lactation, by approximately 80%, compared to that of ovariectomized and intact (diestrous and metestrous cycling) rats while no statistical difference in glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) mRNA level was observed between groups. During the estrous cycle, the level of LHRH-R mRNA in the pituitary was about two-fold higher on diestrous day 2 and the morning of proestrus than that on diestrous day 1 and quickly returned toward control level by noon of proestrus. In addition, we found that GAPDH mRNA levels from a so-called housekeeping gene often thought to be unchanged under different conditions, were significantly higher on proestrus while levels of 18S rRNA were not significantly changed. The large decrease in LHRH-R mRNA during lactation could account for the changes in LHRH binding previously reported.

Animals↗