Effects of macrophage depletion on characteristics of cervix remodeling and pregnancy in CD11b-dtr mice.

To test the hypothesis that macrophages are essential for remodeling the cervix in preparation for birth, pregnant homozygous CD11b-dtr mice were injected with diphtheria toxin (DT) on days 14 and 16 postbreeding. On day 15 postbreeding, macrophages (F4/80+) were depleted in cervix and kidney, but not in liver, ovary, or other non-reproductive tissues in DT-compared to saline-treated dtr mice or wild-type controls given DT or saline. Within 24 h of DT-treatment, the density of cell nuclei and macrophages declined in cervix stroma in dtr mice versus controls, but birefringence of collagen, as an indication of extracellular cross-linked structure, remained unchanged. Only in the cervix of DT-treated dtr mice was an apoptotic morphology evident in macrophages. DT-treatment did not alter the sparse presence or morphology of neutrophils. By day 18 postbreeding, macrophages repopulated the cervix in DT-treated dtr mice so that the numbers were comparable to that in controls. However, at term, evidence of fetal mortality without cervix ripening occurred in most dtr mice given DT-a possible consequence of treatment effects on placental function. These findings suggest that CD11b+ F4/80+ macrophages are important to sustain pregnancy and are required for processes that remodel the cervix in preparation for parturition.

Placental gene expression in a rat 'model' of placental insufficiency.

OBJECTIVE: Placental insufficiency is a major factor associated with pregnancy complications such as miscarriages, intrauterine growth restriction and pre-eclampsia. Recent studies have identified the Brown Norway (BN) rat as a natural 'model' of placental insufficiency associated with decreased trophoblast remodeling of maternal uterine arteries. HYPOTHESIS: Genetic pathways involved in angiogenesis and immune cell regulation are dysregulated in the placenta of BN rats. METHODS: Global gene expression in placentas from BN rats were compared with that from Sprague-Dawley (SD) controls at 17.5 days of gestation using the Affimetrix Rat 1.0 microarray chip, and results confirmed with real-time PCR and immunoblotting. RESULTS: We found significant differences in 272 genes with 108 being up-regulated and 164 down-regulated in BN placentas compared to SD placentas. BN placentas overexpressed genes involved in the renin-angiotensin system (RAS) such as Ace, Ace2, Agtr1a, Nox4, and Ephx2, while key genes involved in angiogenesis, such as Mmp1, Mmp10, Fgfbp1, Esr1, Itga2, Rgs5, and Ccnb1 were down-regulated. We also observed increased expression of Timd2, Itm2a, Irak3, and Csf1r, and decreased expression of Slpi, Ncam1, and Igsf3 in BN placentas. In addition, we observed lower placental weights in BN males compared to BN females, together with increased expression of Cyp1a1 in BN males, as compared to BN females. CONCLUSIONS: The present study demonstrates differential expression of genes involved in blood pressure, angiogenesis and immune cell regulation in BN placenta, and suggests that the RAS may be involved in the pathogenesis of placental dysfunction observed in BN rats.

Elevated total peripheral leukocyte count may identify risk for neurological disability in asphyxiated term neonates.

OBJECTIVE: The present study investigated the relationship between neurologic outcome and total circulating white blood cell (WBC) and absolute neutrophil counts (ANCs) in the first week of life in term infants with hypoxic-ischemic encephalopathy (HIE). STUDY DESIGN: Long-term neurologic outcome at 18 months was measured retrospectively in 30 term neonates with HIE using the Pediatric Cerebral Performance Category Scale (PCPCS) score with outcomes dichotomized as either good or poor. We then compared white blood cell and ANC levels during the first 4 days of life and magnetic resonance imaging (MRI) obtained within the first month life between the two PCPCS groups. MRI was quantified using a validated scoring system. RESULTS: Neonates with good long-term outcomes had significantly lower MRI scores (indicating lesser injury) than neonates with poor outcomes. More importantly, neonates with poor outcomes had significantly higher WBC and ANC levels as early as12 h after birth and up to 96 h after birth compared to those with good outcomes. These data suggest that elevated peripheral neutrophil counts in the first 96 h of life may signal or predict adverse long-term outcome. CONCLUSIONS: Our findings suggest that elevated peripheral neutrophil counts in the first 96 h of life in term infants with HIE may contribute to abnormal neurodevelopmental outcome.

The role of leukocyte traffic and activation in parturition.

OBJECTIVE: This review focuses on the contribution of immune cell trafficking and activities during the initial phase of activation in the process of parturition. Although uterine contractile activity has been the predominant focus for the mechanism that initiates labor, significant cellular and biochemical changes cause remodeling of the cervix well before term. A convergence of evidence suggests that inflammatory processes that involve prostaglandins, nitric oxide, cytokines, as well as systemic and paracrine endocrine mediators may enhance uterine contractility, promote ripening of the cervix, and thus constitute an integrative hypothesis for the initiation of labor. METHODS: Techniques to study the uterus and cervix of pregnant and virgin C3H/HeN mice included light and fluorescence microscopy. Tissues were processed by histochemistry and immunofluorescence. Analytic approaches to enumerate macrophages and assess activation included quantitative stereologic morphometry and laser scanning cytometry. RESULTS: The transition between relative quiescence of the uterus and enhanced contractility involved migration of macrophages from the uterine endometrium and activation of macrophages in the cervix. Before birth, macrophages migrate into the cervix and are activated in the myometrium. CONCLUSION: Immune cell trafficking and activation are part of the initial mechanism that promotes ripening of the cervix, enhances uterine contractility, and initiates parturition. Markers for the conclusion of pregnancy may have diagnostic or therapeutic value to assess the normal progress of labor or identify women at risk of preterm labor.

Photorefractoriness of immune function in male Siberian hamsters (Phodopus sungorus).

Short days induce multiple changes in reproductive and immune function in Siberian hamsters. Short-day reproductive inhibition in this species is regulated by an endogenous timing mechanism; after approximately 20 weeks in short days, neuroendocrine refractoriness to short-day patterns of melatonin develops, triggering spontaneous recrudescence of the reproductive system. It is unknown whether analogous mechanisms control immune function, or if photoperiodic changes in immune function are masked by prevailing photoperiod. In Experiment 1, 3 weeks of exposure to long days was not sufficient to induce long-day-like enhancement of in vitro lymphocyte proliferation in short-day adapted male Siberian hamsters. Experiment 2 tested the hypothesis that immunological photorefractoriness is induced by prolonged exposure to short days. Adult male hamsters were gonadectomized or sham-gonadectomized and housed in long (14 h light/day) or short (10 h light/day) photoperiods for 12, 32 or 40 weeks. Somatic and reproductive regression occurred after 12 weeks in short days, and spontaneous recrudescence was complete after 32-40 weeks in short days, indicative of somatic and reproductive photorefractoriness. In gonad-intact hamsters, 12 weeks of exposure to short days decreased the number of circulating granulocytes and increased the number of B-like lymphocytes. After 32 weeks in short days, these measures were restored to long-day values, indicative of photorefractoriness; castration eliminated these effects of photoperiod. In both intact and castrated hamsters, in vitro proliferation of splenic lymphocytes was inhibited by 12 weeks of exposure to short days. After 40 weeks in short days lymphocyte proliferation was restored to long-day values in intact hamsters, but remained suppressed in castrated hamsters. These results suggest that short-day-induced inhibition of lymphocyte function does not depend on gonadal regression, but that spontaneous recrudescence of this measure is dependent on gonadal recrudescence. In Experiment 3, in vitro treatment with melatonin enhanced basal proliferation of lymphocytes from male hamsters exposed to short days for 12 weeks, but had no effect on lymphocytes of photorefractory hamsters or long-day control hamsters. Lymphocytes of castrated hamsters were unresponsive to in vitro melatonin, suggesting that photoperiodic changes in gonadal hormone secretion may be required to activate mechanisms which permit differential responsiveness to melatonin depending on phase in the annual reproductive cycle. Together, these data indicate that, similar to the reproductive system, the immune system of male Siberian hamsters exhibits refractoriness to short days.

Photoperiod, reproduction, and immunity in select strains of inbred mice.

The purpose of this study was to determine whether decreased day lengths affect reproduction or the immune system in inbred mice. Irrespective of a nocturnal pineal melatonin rise, the signal for day length information, body and testis weights were the same in various strains 8 weeks after transfer from long to short days (16 to 8 h of light/day) compared to mice that remained in long days. Serum testosterone was unaffected by the photoperiod shift. The second goal was to determine whether the shift from long to short days influenced lymphocyte populations in spleen or blood, as well as innate and cell-mediated immune cell functions in C3H/HeN mice, an inbred strain with a robust melatonin rhythm. By flow cytometry, a stable percentage and number of B cells, T cells, and natural killer cells were identified in spleen from mice in both long and short days during the day and night. This complement of immunophenotypes in spleen suggests that equivalent functional capabilities persist in secondary lymphoid tissue of mice irrespective of day length. This was supported by findings that cytolytic activity by splenic natural killer cells (innate immunity) and antigen-induced T cell-dependent B cell antibody production (adaptive immunity) were similar in mice in long and short days. In blood, cell numbers but not helper T cell subset percentages (i.e., naive, memory, cytotoxic, or activated) were augmented in mice in short compared to long days, a consequence of increased circulating B cells. Day length differences in certain immunophenotypes in circulation may forecast photoperiod-mediated alterations in responsiveness to pathogens that are associated with a change in season. At night, the reduced proportion of cytotoxic T cells (long and short days), as well as increases in the percentage of activated T cells (long days), B cells (short days), and NK cell activity (long and short days) relative to daytime, suggests that surveillance and function by select immunophenotypes may adapt to circadian transitions even in highly inbred species. Thus, inbred mice retain capabilities for photoperiod to influence trait-specific aspects of immune cell but not reproductive function.

Aging-dependent changes in the effect of daily melatonin supplementation on rat metabolic and behavioral responses.

Pineal melatonin secretion has been reported to commonly decrease with aging, whereas intra-abdominal adiposity, plasma insulin and plasma leptin levels tend to increase. We recently demonstrated that daily melatonin administration starting at middle age suppressed male rat intra-abdominal fat, plasma leptin and plasma insulin to youthful levels, suggesting that aging-related changes in pineal melatonin secretion and in energy regulation may be functionally related. Accordingly, we have now investigated the effects of daily melatonin treatment on energy regulation in young versus middle-aged male Sprague Dawley rats. Addition of melatonin to the drinking water (0.2 microg/mL) produced nocturnal and diurnal plasma melatonin concentrations in middle-aged rats (12 months) equivalent to those of young adult (5 months) rats. Administration of this melatonin dosage every day for 10 wk starting at 10 months of age suppressed (P < 0.01) relative intra-abdominal fat, non-fasted plasma insulin and plasma leptin by 27, 39, and 51%, respectively (vs. vehicle-treated controls). In contrast, administration of melatonin for 10 wk starting at 3 months of age did not significantly alter (P> 0.10) any of these parameters. The melatonin administration stimulated (102%, P < 0.001) behavioral responsiveness of the middle-aged rats in a test of response to novelty, restoring youthful levels, but did not significantly alter behavioral responsiveness of the young rats. These results suggest that suppression of intra-abdominal adiposity and plasma leptin and insulin levels and stimulation of behavioral responsiveness in response to daily exogenous melatonin begins at middle age, coincident with and likely dependent upon the aging-associated decline in endogenous pineal melatonin secretion. These results further suggest that appropriate melatonin supplementation may potentially provide therapy or prophylaxis not only for the insulin resistance, increased intra-abdominal fat and resulting pathologies that occur with aging, but also for some aging-associated behavioral changes.

Photoperiod modulates the inhibitory effect of in vitro melatonin on lymphocyte proliferation in female Siberian hamsters.

In Siberian hamsters (Phodopus sungorus), short days suppress reproductive function and lymphocyte proliferation. To determine whether melatonin influences cell-mediated immunity through a direct action on lymphocyte proliferation, in vitro responsiveness to mitogens and melatonin was assessed in systemic and splenic lymphocytes from adult female Siberian hamsters housed in either long or short days for 13 weeks. Short days provoked reproductive regression and reduced lymphocyte proliferation. Physiological concentrations of melatonin (50 pg/ml) inhibited in vitro proliferation of circulating lymphocytes, whereas higher concentrations (> or = 500 pg/ml) were required to inhibit proliferation of splenic lymphocytes. Immunomodulatory effects of melatonin were restricted to lymphocytes from long-day hamsters-in vitro melatonin had no effect on circulating or splenic lymphocytes from females in short days. Responsiveness to melatonin in short-day lymphocytes may be restrained by the already expanded nightly pattern of melatonin secretion in short days. These data support the hypothesis that melatonin acts directly on lymphocytes from long-day hamsters to suppress blastogenesis.

Aging, reproduction, and the melatonin rhythm in the Siberian hamster.

The present study tested the hypothesis that responsiveness to melatonin, the presence of the melatonin rhythm in circulation, and parameters of the GnRH neuron system are sustained across the aging continuum in Siberian hamsters. Afternoon melatonin injections induced testicular atrophy in 42% of aged males compared with 100% of adult males. The proportion of aged males failing to respond to the melatonin injections was similar to the proportion that failed to undergo testicular regression upon exposure to short days. Exposure to short days induced testicular atrophy in juvenile and adult hamsters; however, regression was incomplete or absent in 43% of aged males. The nocturnal rise in melatonin was similar with regard to duration and peak amplitude, and appropriate with respect to photoperiod in 25-day-old juveniles, adult (5 months), and aged (17 months) hamsters. Neither advanced age nor timed melatonin treatments affected GnRH neuron numbers or distribution. Fertility was maintained in aged and adult males to a comparable extent with respect to latency to first litter and number of pups per litter; reproductive success was dramatically reduced in aged compared with adult females. Because melatonin rhythms accurately reflect day length information throughout the continuum from puberty to advanced age, the present evidence suggests that limitations in testis regression in response to short days or exogenous melatonin in a subset of aged males result from a reduced ability to respond to melatonin. In the wild, failure to undergo testicular regression in the presence of shortening day lengths may extend the breeding season of aged males.

Temporal reorganization of the suprachiasmatic nuclei in hamsters with split circadian rhythms.

A dual oscillator basis for mammalian circadian rhythms is suggested by the splitting of activity rhythms into two components in constant light and by the photoperiodic control of pineal melatonin secretion and phase-resetting effects of light. Because splitting and photoperiodism depend on incompatible environmental conditions, however, these literatures have remained distinct. The refinement of a procedure for splitting hamster rhythms in a 24-h light-dark:light-dark cycle has enabled the authors to assess the ability of each of two circadian oscillators to initiate melatonin secretion and to respond to light pulses with behavioral phase shifting and induction of Fos-immunoreactivity in the suprachiasmatic nuclei (SCN). Hamsters exposed to a regimen of afternoon novel wheel running (NWR) split their circadian rhythms into two distinct components, dividing their activity between the latter half of the night and the afternoon dark period previously associated with NWR. Plasma melatonin concentrations were elevated during both activity bouts of split hamsters but were not elevated during the afternoon period in unsplit controls. Light pulses delivered during either the nighttime or afternoon activity bout caused that activity component to phase-delay on subsequent days and induced robust expression of Fos-immunoreactivity in the SCN. Light pulses during intervening periods of locomotor inactivity were ineffective. The authors propose that NWR splits the circadian pacemaker into two distinct oscillatory components separated by approximately 180 degrees, with each expressing a short subjective night.

Reproductive, neuroendocrine, and immune consequences of acute exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin in the Siberian hamster.

The present study tested the hypothesis that acute treatment with 2, 3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) impairs fertility, disrupts the nocturnal melatonin rhythm, and suppresses lymphocyte function. Adult Siberian hamsters administered 2 or 100 microg TCDD/kg body weight/0.2 ml sesame oil had a delayed latency to first litter and an increased adult mortality compared to hamsters given 0.1 microg/kg or vehicle. Within 75 days of TCDD treatment, full reproductive capabilities were achieved. Moreover, the nocturnal melatonin rhythm was not disrupted in adults administered TCDD or in their progeny. Lymphocyte activity varied with respect to time of day and treatment. Lymphocyte proliferation was enhanced at night irrespective of TCDD treatment; during the day, 2 wk after the 2-microg/kg treatment, blastogenesis was reduced compared to that in the 0.1-microg/kg group or in vehicle-treated controls. In contrast, TCDD did not affect the mixed lymphocyte reaction in response to allogeneic antigen when assessed at 2 and 20 wk post-treatment. Thus, findings indicate that TCDD produced acute effects on fertility, mortality, and systemic lymphocyte proliferation, but long-lasting effects on specific aspects of reproductive, neuroendocrine, and immune cell functions were not observed.

Distribution and activation of uterine mononuclear phagocytes in peripartum endometrium and myometrium of the mouse.

The present study tested the hypothesis that macrophage distribution and activation are enhanced in the uterus before term. Mid-uterine horn tissue strips from mice on Days 15 and 18 of pregnancy, the day of birth (= Day 19), and one day postpartum were paraffin-embedded and then sectioned, stained with a monoclonal pan-macrophage marker (BM8), and processed for visualization and quantification of resident macrophages per nuclear area. Macrophages were dispersed throughout the endometrium and subluminal epithelium; cell numbers declined on the day before term, then increased postpartum. Within myometrium, macrophages congregated in stroma surrounding muscle bundles, and staining was enhanced near term. Macrophage numbers were similar in pregnant and postpartum uteri, enhanced more than 2-fold over those in nonpregnant controls. Uterine sections were also analyzed by laser-scanning cytometry to enumerate activated macrophages (i.e., those that express the intercellular adhesion molecule marker CD54+) and to determine cell cycle (propidium iodide fluorescence). Activated macrophages were directly proportional to cell numbers and, by cell cycle analysis, were not terminally differentiated. Highest cell numbers occurred on Day 15: 4-fold greater than those in nonpregnant controls and 2-fold higher than those at Day 18 or in postpartum groups. These findings indicate a decline in endometrial macrophage numbers at least one day before the onset of parturition and raise the possibility that trafficking of this immune cell may contribute to onset of labor.

Photic entrainment of circannual rhythms in golden-mantled ground squirrels: role of the pineal gland.

Entrainment of circannual rhythms of body mass and reproduction was monitored for 3 years in female golden-mantled ground squirrels maintained in a simulated natural photoperiod. Both pinealectomized and pineal-intact squirrels generated circannual rhythms of body mass and estrus, but only the intact animals entrained these rhythms to a period of 365 days. In the second and third years after treatment, the period of the body mass rhythm was significantly shorter than 365 days for pinealectomized squirrels, and variance in tau among these animals was significantly greater than for intact squirrels. A similar pattern was evident in the rhythm of reproduction, which was phase-disrupted in pinealectomized squirrels but entrained in intacts. Seasonal changes in duration of nocturnal melatonin secretion by the pineal appear to be necessary to produce phase-delays required to entrain the circannual clock to a period of 12 months.

In vitro melatonin treatment enhances splenocyte proliferation in prairie voles.

The seasonal effects of photoperiod on reproduction are mediated by melatonin, and it is hypothesized that increased immune function in short days is due to the increase in the duration of nightly melatonin secretion. Melatonin can act both directly and indirectly on target tissue within the immune system. The present study sought to tease apart the direct and indirect effects of melatonin on one aspect of immune function by examining the influence of in vitro melatonin on splenocyte proliferation in female prairie voles held in long (LD 16:8) or short (LD 8:16) days. Splenocyte proliferation in response to the T-cell mitogen concanavalin A was enhanced by the addition of melatonin in vitro, as compared to cultures receiving no melatonin. Body mass increased in short-day housed prairie voles, indicating that the animals were responsive to photoperiod. However, photoperiod did not affect splenocyte proliferation in the present study. These results support the hypothesis that melatonin exerts a direct effect on splenocyte proliferation, potentially via high-affinity melatonin receptors localized on splenocytes. The findings also indicate that, irrespective of photoperiod, melatonin exerts direct effects on splenocytes to enhance immune function.

Daily melatonin administration to middle-aged male rats suppresses body weight, intraabdominal adiposity, and plasma leptin and insulin independent of food intake and total body fat.

Pineal melatonin secretion declines with aging, whereas visceral fat, plasma insulin, and plasma leptin tend to increase. We have previously demonstrated that daily melatonin administration at middle age suppressed male rat intraabdominal visceral fat, plasma leptin, and plasma insulin to youthful levels; the current study was designed to begin investigating mechanisms that mediate these responses. Melatonin (0.4 microg/ml) or vehicle was administered in the drinking water of 10-month-old male Sprague Dawley rats (18/treatment) for 12 weeks. Half (9/treatment) were then killed, and the other half were submitted to cross-over treatment for an additional 12 weeks. Twelve weeks of melatonin treatment decreased (P<0.05) body weight (BW; by 7% relative to controls), relative intraabdominal adiposity (by 16%), plasma leptin (by 33%), and plasma insulin (by 25%) while increasing (P<0.05) locomotor activity (by 19%), core body temperature (by 0.5 C), and morning plasma corticosterone (by 154%), restoring each of these parameters toward more youthful levels. Food intake and total body fat were not changed by melatonin treatment. Melatonin-treated rats that were then crossed over to control treatment for a further 12 weeks gained BW, whereas control rats that were crossed to melatonin treatment lost BW, but food intake did not change in either group. Feed efficiency (grams of BW change per g cumulative food intake), a measure of metabolic function, was negative in melatonin-treated rats and positive in control rats before cross-over (P<0.001); this relationship was reversed after cross-over (P<0.001). Thus, melatonin treatment in middle age decreased BW, intraabdominal adiposity, plasma insulin, and plasma leptin, without altering food intake or total adiposity. These results suggest that the decrease in endogenous melatonin with aging may alter metabolism and physical activity, resulting in increased BW, visceral adiposity, and associated detrimental metabolic consequences.

Maturation of lymphocyte immunophenotypes and memory T helper cell differentiation during development in mice.

The goal of this study was to systematically investigate the ontogeny of lymphoid populations throughout postnatal development. In CD-1 mice, peak lymphocyte numbers occurred in blood on postnatal day 10 (d10) including those for natural killers (NK1.1), B cells (CD19), T helper (CD3CD4), naïve T helper (CD4CD62LposCD44low), memory T helper (CD4CD62LnegCD44high), and T cytotoxic (CD3CD8) cells. As percent of total lymphocytes, peaks were achieved by d10 for all T helper subtypes but not B cells which declined to a nadir. In spleen, lymphocyte numbers increased exponentially after d10. Proportionately, NK and T cells peaked on d10, declined by d20, and increased 2-3-fold by d45. Naive T cells constituted the majority of lymphocytes during development while memory cells gained to 2.2% (blood) and 12% (spleen) by d20. C57BL/6 mice had similar profiles except that the B cell nadir and T cell subset peaks were at d5. Peripheralization of critical numbers of lymphocytes by d10, and importantly, development of a repertoire of memory cells by d20, may define immune response capabilities that close the period of immaturity for the neonate.

Role of photoperiod and the pineal gland in T cell-dependent humoral immune reactivity in the Siberian hamster.

The present study tested the hypothesis that antibody production in response to xenoantigen is modulated by daylength and dependent upon the pineal gland. Alter injection of sheep erythrocytes (SRBC), serum immunoglobulin (Ig) concentrations were 5-fold lower in hamsters in short versus long days. Pinealectomy (Pinx) abolished the nocturnal melatonin rhythm, blocked short-day-mediated testis regression, and eliminated the short-day reduction in Ig production after SRBC treatment. Antibody titers in response to SRBC were equivalently augmented in short-day Pinx and long-day sham hamsters. The results indicate that photoperiodic effects on T cell-dependent humoral immunity are dependent upon the pineal gland. These findings raise the possibility that day length-associated changes in some immune system functions are mediated by the pineal melatonin rhythm.

Ventromedial hypothalamic mediation of photoperiodic gonadal responses in male Syrian hamsters.

Short day lengths induce testicular regression in seasonally breeding Syrian hamsters. To test whether the ventromedial hypothalamus is necessary to maintain reproductive quiescence once testicular regression has been achieved, photoregressed male hamsters were subjected to lesions of the ventromedial hypothalamus (VMHx), pinealectomy (Pinx), or sham operation (Sham). VMHx hamsters underwent accelerated gonadal recrudescence compared to Pinx and Sham hamsters. Recovery of prolactin concentrations (PRL) to values characteristic of long-day hamsters was hastened in the VMHx animals compared to Sham hamsters. Concentrations of follicle stimulating hormone (FSH) increased prematurely in both the VMHx and Pinx animals, beginning a few weeks after surgery. By the time the gonads had undergone recrudescence and the hamsters were refractory to melatonin, PRL and FSH concentrations had returned to baseline long-day values in all groups; there was no evidence of hypersecretion of either hormone in any of the animals with lesions. Melatonin concentrations of VMHx hamsters did not differ from those of sham-operated animals, but because only a single determination was made, it remains possible that VMH damage altered the duration of nightly melatonin secretion. An intact VMH appears to be essential for the continued maintenance of reproductive suppression induced by exposure to short day lengths; these and earlier findings suggest that the VMH-dorsomedial hypothalamic complex mediates regression of the reproductive apparatus during decreasing day lengths of late summer and early autumn and also is necessary to sustain regression during the winter months.

Maturation of spontaneous and agonist-induced uterine contractions in the peripartum mouse uterus.

This study tested the hypothesis that the uterus achieves maximum contractile capabilities before the onset of labor. Basal and agonist-stimulated contractions were assessed in uterine strips on Day 15 or 18 of pregnancy, the day of parturition, or 1 day postpartum (n = 4-13 per group). Spontaneous contractions were evident in all groups (n = 4-13 per gestational group); contraction frequency was greater in peripartum groups than in virgin controls ( approximately 4.6 versus 2.8/200 sec). Peak amplitude was nearly 9-fold higher on Days 15 and 18 and over 30-fold higher in the postpartum and 1 day postpartum groups than in nonpregnant mice. Maximum frequency and peak amplitude were achieved in response to 10(-6) to 10(-8) M oxytocin or arginine vasopressin (OT(max) or AVP(max)). Frequency of contractions in response to OT(max) peaked on Day 18 and then declined. Contraction amplitude increased 5-fold on Day 15, declined on the day of birth (equivalent to nonpregnant level), then rebounded to peak on postpartum Day 1. AVP(max) similarly increased frequency and amplitude of contractions, except that maximum contraction amplitude occurred postpartum. Thus, an endogenous oscillator, residing in the uterus, sustains high basal and agonist-induced contraction frequency during pregnancy. Although acceleration of this pacemaker occurred before term, the data suggest that peripartum increases in contraction amplitude characterize the transition to the powerful synchronous contractions of parturition.

Macrophage trafficking in the uterus and cervix precedes parturition in the mouse.

Immune activation is implicated in the etiology of preterm labor, but little is known about macrophage number or distribution in the uterus or cervix at term. This study tested the hypothesis that macrophages migrate into the reproductive tract before the onset of parturition. Paraffin-embedded sections from the mid-uterine horn and cervix of C3/HeN mice on Days 15 and 18 of pregnancy, the day of birth (Day 19), and 1 day postpartum were stained with a pan-macrophage marker to analyze cell numbers and distribution. During pregnancy, uterine macrophages were dispersed in endometrium, usually associated with vasculature and subluminal epithelium. In myometrium, macrophages were clustered in stromal connective tissue; near term and postpartum, cells appeared to surround the muscle bundles. Total macrophage numbers were increased on Day 15 relative to those in nonpregnant controls, declined before birth, and increased postpartum. In the cervix, macrophages congregated in subepithelium, often perivascular or near ganglia. Macrophage numbers in the cervix peaked on Day 18, then declined to nonpregnant levels by the day after birth. Thus, macrophage numbers in the uterus were inversely related to those in the cervix. These findings raise the possibility that macrophages and their products may be involved in uterine contractility and cervical remodeling during the processes of parturition.