Specific neurotoxin lesions of median raphe serotonergic neurons disrupt maternal behavior in the lactating rat.

Impairments in lactation after electrolytic lesions of the median raphe (MR) nucleus have been corrected by treatment with PRL. Specific serotonin neurotoxin lesions were used in the present study to determine whether decrements in litter growth after electrolytic lesions could be attributed to serotonergic neuron damage at the MR locus, and whether MR lesions (MRL) disrupted suckling-induced PRL release. Intracerebral microinjection of 5,7-dihydroxytryptamine (5,7-DHT) into the MR nucleus produced dose-related decrements in litter growth after either 4 micrograms (sham, 1.35 +/- 0.05; MRL, 1.04 +/- 0.05 g/pup X day; P less than 0.001) or 8 micrograms 5,7-DHT (sham, 1.35 +/- 0.06; MRL, 0.87 +/- 0.11 g/pup X day; P less than 0.001). Despite hypothalamic serotonin depletions of 15% and 55%, respectively, for the two doses of 5,7-DHT, there was no difference between sham and MRL animals in either basal or suckling-induced PRL release. When lesions were placed on day 1 of lactation (L) so that killing on day 7-L corresponded to the early maximal neurotoxin effect, MRL mothers still showed litter growth decrements (0.37 +/- 0.07; sham, 0.98 +/- 0.08 g/pup X day; P less than 0.001) and normal PRL values. When maternal behavior was examined, MRL animals exhibited a higher incidence of abnormal behaviors (failure to retrieve pups, cannibalism, and failure to initiate suckling during a 1-h test period; Fisher's exact P, Sham vs. MRL, less than 0.01, less than 0.05, and 0.15, respectively) than sham animals or animals with 5,7-DHT lesions in the dorsal raphe nucleus or superior colliculus. In addition, suckling behavior scores, determined from daily suckling behavior observations, were lowest in the MRL group and correlated with litter growth only in this group (r = 0.789; P less than 0.01). These data suggest that serotonergic elements in the MR nucleus play an obligatory role in maintaining normal maternal behavior during lactation, but they are not involved in suckling induced PRL release.

Dorsal raphe-hypothalamic projections provide the stimulatory serotonergic input to suckling-induced prolactin release.

The present study was carried out to determine whether dorsal raphe serotonergic neurons are involved in the regulation of suckling-induced PRL release. Neurotoxin lesions were placed stereotaxically in lactating rats on day 1 of lactation by 5,7-dihydroxytryptamine microinjection into the dorsal raphe (DR), median raphe (MR), or superior colliculus (SC), an area devoid of serotonergic perikarya. Litters were adjusted to eight pups each and weighed daily to determine litter growth rates. On day 7 of lactation, litters were separated from mothers for 8 h, after which six healthy foster pups were provided for a 30-min suckling stimulus. Animals were killed by decapitation immediately after suckling, plasma was collected for RIA of PRL, and brains were frozen and dissected for determination of hypothalamic, caudate, and hippocampal serotonin (5-HT) using the enzymatic-isotopic assay procedure. Litter growth rates from days 1-7 of lactation were significantly different among lesion groups (P less than 0.005), with litters from SC-lesioned animals (SCL) growing similarly to the sham group (sham, 0.924; SCL, 0.941 g/pup . day). In contrast, growth rates of litters from both DR-lesioned (DRL) and MR-lesioned (MRL) animals were significantly depressed (DRL, 0.596; MRL, 0.449 g/pup . day; P less than 0.05 and P less than 0.01, respectively). 5-HT levels in hypothalamus, caudate nuclei, and hippocampus were similar in the sham and SCL groups, whereas hypothalamic 5-HT was depleted by 63% and 55%, respectively, in the DRL and MRL groups. Despite impairments in growth rate and litter survival in both the DRL and MRL groups, only DRL animals showed significant decrements in suckling-induced PRL release (DRL, 288 +/- 107; sham, 837 +/- 134 ng NIAMDD rat PRL RP-1/ml; P less than 0.05) after 5-HT-depleting lesions. The results suggest a specificity of function within the raphe system during lactation; DR 5-HT neurons which project to the hypothalamus provide stimulatory inputs to suckling-induced PRL release, whereas MR 5-HT neurons influence litter growth and survival via their role in maternal behavior.

Time course of the luteinizing hormone surge in cannulated rats: quantitative variance estimates within individual rats over successive cycles.

The present study was designed to compare quantitative variance estimates in the profile of the luteinizing hormone (LH) surge within individual rats over successive proestrous (PE) days (WITHIN) with the variability between rats (BETWEEN). Sprague-Dawley female rats were implanted under ether anesthesia with indwelling intracardiac cannulas. On successive PE afternoons of normal 4-day cycles, hourly blood samples (0.25 ml) were collected via the cannula from 1400-2000 h for radioimmunoassay of plasma LH. Three characteristics which reflected the profile of the LH surge were examined: the time of onset of LH release, the time of peak LH release, and the magnitude of peak LH release. Twenty-one animals yielded LH surge data on a total of 42 PE days with a mean (+/- SD) time of onset = 1534 h +/- 66 min, time of peak = 1730 h +/- 75 min, and magnitude of peak = 1176 +/- 441 ng NIAMDD-Rat LH-RP-1/ml plasma. Variance estimates BETWEEN and WITHIN animals were determined by analysis of variance and the method of Vaughan and Corballis (1969) for calculating percent of total variance. Differences BETWEEN in time of onset of LH release approached significance (P = 0.05-0.10) and contributed 50.5% of the total variance compared to a negative value for differences WITHIN. Differences BETWEEN in time of peak LH release were significant (P less than 0.05) and contributed 51.5% of total variance compared to a negative value for differences WITHIN. In contrast, for the magnitude of peak LH release, neither differences BETWEEN nor WITHIN contributed substantially to total variance (both negative values), with the major contribution from the residual term.(ABSTRACT TRUNCATED AT 250 WORDS)

Median raphe stimulation and sham procedures inhibit the LH surge.

Acute electrolytic stimulation of the median raphe (MR) nucleus (MR Stim) between 13:00 and 16:00 h on proestrus blocked ovulation in 10 of the 21 rats tested, whereas all sham-treated (MR Sham, no current) animals ovulated normally (n = 12). MR Stim at 16:00 h on proestrus in individual cannulated rats 'turned off' the luteinizing hormone (LH) surge in all animals tested. Unexpectedly, MR Sham treatment also abruptly inhibited the LH surge, although all animals ovulated. Manipulations associated with acute surgical procedures (puncture of the dura mater, ether anesthesia alone) were also found to inhibit the LH surge, with the magnitude of LH inhibition directly related to the severity of the operative procedures performed (p less than 0.025). The results suggest (1) that inhibition of ovulation following MR stimulation can be attributed to a response to stress qualitatively similar but more severe than that experienced by the MR sham group, and (2) that inhibitory inputs to the ovulatory LH release mechanism cannot be studied using acute surgical procedures.

Impairments in lactation in the rat following destruction of the median raphe nucleus.

Lesions of the median raphe (MR) nucleus were placed in cycling female rats and their ability to lactate was evaluated following subsequent pregnancies. Pups from MR-lesioned (MRL) animals grew more slowly and had greatly impaired survival rates compared to pups from sham-lesioned animals. Chronic treatment of MRL mothers with oxytocin (Oxy; 1 IU, s.c., once or twice/day) did not increase the growth rates of their litters. Acute responses to exogenous Oxy (1 IU, i.p.) in MRL mothers, measured by the weight gain of litters during 1/2-h suckling intervals before and after injection, were marginally significant. Milk yield during the total hour suckling period (stomach contents of pups) was clearly less in the MRL animals (p less than 0.01). Treatment with either prolactin (Prl; 250 microgram, twice/day), Prl + GTC (4 mg/kg gorwth hormore, 30 microgram/kg thyroxine, 0.5 mg/rat cortisol, once/day), or 5-HTP (75 mg 5-hydroxytryptophan/kg, twice/day) did not improve the growth rates of litters from MRL animals. However, when milk yield (stomach contents after 1 h) following a 14-h non-suckling interval was measured, lactogenic hormones (Prl or Prl + GTC) restored milk yield in MRL animals to control levels. This response was clearly not dependent upon exogenous Oxy. These results suggest that deficits in the release of lactogenic hormones are involved in the impairments in lactation following lesions of the MR nucleus.

Patterns of LH and FSH release from perifused rat pituitaries in response to infusions of hypothalamic extract.

A continuous flow incubation (perufusion) system was developed in which the secretory responses of pools of hemipituitaries from adult male rats to hypothalamic extract (HE) were characterized by serial radioimmunoassay of LH and FSH in the effluent medium. There was in initial massive release of LH and FSH which, in the absence of HE, declined to low basal levels at a rate which depended on the flow rate. Thereafter, the baseline for LH continued to decline gradually while that for FSH was stable. The rate of LH and FSH release rose abruptly after addition of the HE to the medium and returned promptly to baseline after withdrawal of the HE. During continuous infusion of HE for five hours, LH secretion was maintained at a relatively constant, elevated level. The responses to repeated identical pulses of HE were highly reproducible. The variability between responses by any one pool of tissue was significantly less than between responses of separate pituitary pools including pools comprising right and left halves of the same glands. For any given pool of pituitaries of the relationships were linear between: 1) duration of HE pulses (concentration constant) and increases in LH output, and 2) log of concentration of HE (pulse duration constant) and increases inLH and in FSH output. Consistent responses were obtained for up to 12 hr, the maximum period tested.