Gland Wet Weight Research Articles

Synergistic streptococcal phage λSA2 and B30 endolysins kill streptococci in cow milk and in a mouse model of mastitis.

Bovine mastitis results in billion dollar losses annually in the USA alone. Streptococci are among the most relevant causative agents of this disease. Conventional antibiotic therapy is often unsuccessful and contributes to development of antibiotic resistance. Bacteriophage endolysins represent a new class of antimicrobials against these bacteria. In this work, we characterized the endolysins (lysins) of the streptococcal phages λSA2 and B30 and evaluated their potential as anti-mastitis agents. When tested in vitro against live streptococci, both enzymes exhibited near-optimum lytic activities at ionic strengths, pH, and Ca(2+) concentrations consistent with cow milk. When tested in combination in a checkerboard assay, the lysins were found to exhibit strong synergy. The λSA2 lysin displayed high activity in milk against Streptococcus dysgalactiae (reduction of CFU/ml by 3.5 log units at 100 μg/ml), Streptococcus agalactiae (2 log), and Streptococcus uberis (4 log), whereas the B30 lysin was less effective. In a mouse model of bovine mastitis, both enzymes significantly reduced intramammary concentrations of all three streptococcal species (except for B30 vs. S. dysgalactiae), and the effects on mammary gland wet weights and TNFα concentrations were consistent with these findings. Unexpectedly, the synergistic effect determined for the two enzymes in vitro was not observed in the mouse model. Overall, our results illustrate the potential of endolysins for treatment of Streptococcus-induced bovine mastitis.

Tracheal occlusion conditioning causes stress, anxiety and neural state changes in conscious rats

Evidence from human and animal studies indicates that mechanical loads to breathing are stressful stimuli and evoke compensatory behaviours. Conditioning of stressful stimuli is known to cause changes in basal stress levels and behaviour. Individuals with respiratory obstructive diseases repeatedly experience bouts of airway obstruction, which may act as a form of conditioning, and often have affective disorders, such as anxiety and depression. It is unknown whether the development of affective disorders in these individuals results from the unexpected recurring respiratory perturbations. To investigate this possibility, we developed a model to elicit tracheal occlusion (TO) in conscious rats and exposed them to 10 days of TO conditioning. We hypothesized that healthy, conscious animals exposed to TO conditioning would develop stress and anxiety and would have modulated neural activity in respiratory, stress, discriminative and affective neural regions. Following TO conditioning, rats had increased basal corticosterone levels, greater adrenal weights and elevated anxiety levels compared with animals not receiving TO. Significant increases in cytochrome oxidase staining were found in brainstem respiratory nuclei, periaqueductal grey, dorsal raphe, thalamus and insular cortex. These results suggest that healthy animals develop stress and anxiety responses to respiratory load conditioning via inescapable tracheal occlusions, which may be mediated through state changes in specific brain nuclei.

Chimeric Phage Lysins Act Synergistically with Lysostaphin To Kill Mastitis-Causing Staphylococcus aureus in Murine Mammary Glands

Staphylococci cause bovine mastitis, with Staphylococcus aureus being responsible for the majority of the mastitis-based losses to the dairy industry (up to $2 billion/annum). Treatment is primarily with antibiotics, which are often ineffective and potentially contribute to resistance development. Bacteriophage endolysins (peptidoglycan hydrolases) present a promising source of alternative antimicrobials. Here we evaluated two fusion proteins consisting of the streptococcal λSA2 endolysin endopeptidase domain fused to staphylococcal cell wall binding domains from either lysostaphin (λSA2-E-Lyso-SH3b) or the staphylococcal phage K endolysin, LysK (λSA2-E-LysK-SH3b). We demonstrate killing of 16 different S. aureus mastitis isolates, including penicillin-resistant strains, by both constructs. At 100 μg/ml in processed cow milk, λSA2-E-Lyso-SH3b and λSA2-E-LysK-SH3b reduced the S. aureus bacterial load by 3 and 1 log units within 3 h, respectively, compared to a buffer control. In contrast to λSA2-E-Lyso-SH3b, however, λSA2-E-LysK-SH3b permitted regrowth of the pathogen after 1 h. In a mouse model of mastitis, infusion of 25 μg of λSA2-E-Lyso-SH3b or λSA2-E-LysK-SH3b into mammary glands reduced S. aureus CFU by 0.63 or 0.81 log units, compared to >2 log for lysostaphin. Both chimeras were synergistic with lysostaphin against S. aureus in plate lysis checkerboard assays. When tested in combination in mice, λSA2-E-LysK-SH3b and lysostaphin (12.5 μg each/gland) caused a 3.36-log decrease in CFU. Furthermore, most protein treatments reduced gland wet weights and intramammary tumor necrosis factor alpha (TNF-α) concentrations, which serve as indicators of inflammation. Overall, our animal model results demonstrate the potential of fusion peptidoglycan hydrolases as antimicrobials for the treatment of S. aureus-induced mastitis.

Quantification of mammary gland tissue size and composition changes after weaning in sows1

The objectives of this study were to characterize the tissue compositional changes in porcine mammary glands after weaning and to determine whether administration of estradiol alters the profile of these tissue changes. Forty-five primiparous sows were assigned randomly to one of two treatment groups after weaning, control or estrogen treated. Estrogen-treated sows received twice-daily injections of estradiol-17beta (0.125 mg/kg of BW); control sows received vehicle injections. Sows were weaned at d 21 of lactation and killed on either d 0 (d of weaning; n = 5) or on d 2, 3, 4, 5, or 7 after weaning (n = 4 per treatment on each day). Teat order relative to suckling behavior was observed on the day before weaning to determine which mammary glands the piglets suckled. Suckled and non-suckled glands were identified from the teat order observation, and individual mammary glands were collected at slaughter. Mammary glands were trimmed of skin and extraneous fat pad, individually weighed, and bisected to measure cross-sectional area. The remaining half of each gland was ground and stored at -20 degrees C for chemical analyses. Frozen tissue was used for measuring tissue DNA, DM, protein, fat, and ash contents. Suckled mammary glands of sows undergo significant and dramatic changes during the initial 7 d after weaning, with significant changes occurring even by d 2 after weaning. Mean cross-sectional area of parenchymal tissue in suckled mammary glands decreased from 59.7 +/- 2.1 cm2 on the day of weaning to 26.8 +/- 2.3 cm2 by d 7 after weaning (P < 0.0001). Mammary gland wet weight decreased from 485.9 +/- 22.0 g on the day of weaning to 151.5 +/- 24.8 g by d 7 after weaning (P < 0.0001), whereas DNA decreased from 838.8 +/- 46.2 g on the day of weaning to 278.4 +/- 52.5 g by d 7 after weaning (P < 0.0001). The changes in gland wet weight and DNA during the period of mammary gland involution in the sow represent loses of over two-thirds of the parenchymal mass and nearly two-thirds of the cells that were present on the day of weaning. Estrogen treatment did not affect overall mammary involution during the first 7 d after weaning. Mammary glands that were not suckled during lactation had no further loss of parenchymal tissue during the first 7 d after weaning. Mammary gland involution in the sow is a rapid process and is probably irreversible within 2 or 3 d after weaning.

Inability of overexpressed des(1-3)human insulin-like growth factor I (IGF-I) to inhibit forced mammary gland involution is associated with decreased expression of IGF signaling molecules.

Overexpression of des(1-3) human insulin-like growth factor I (IGF-I) in the mammary glands of transgenic mice (WAP-DES) inhibits apoptosis during natural, but not forced, mammary involution. We hypothesized that this differential response would correlate with the expression of IGF signal transducers. Forced and natural involution were analyzed in nontransgenic and WAP-DES mice beginning on day 16 postpartum. During natural involution, mammary gland wet weight was higher and apoptosis was lower in WAP-DES than in nontransgenic mice. The WAP-DES transgene had no effect on these parameters during forced involution. Mammary tissue concentrations of the transgene protein were 2- to 10-fold higher than those of endogenous IGF-I. Western blot analysis of pooled mammary tissue extracts demonstrated only slightly higher phosphorylation of the IGF signal transducers insulin receptor substrate-1 (IRS-1) and Akt in the WAP-DES than in nontransgenic mice. Dramatic early reductions in phospho-IRS-1, phospho-Akt, IRS-1, IRS-2, and Akt proteins occurred during forced, but not natural, involution. The abundance of the IGF-I receptor and the messenger RNAs for the IGF-I receptors, IRS-1 and -2, were not affected by either genotype or involution. These findings support the conclusions that mammary cells lose their responsiveness to insulin-like signals during forced involution, and that posttranscriptional or posttranslational regulation of IRS-1 and IRS-2 may play a role in this loss.

Changes in tissue composition associated with mammary gland growth during lactation in sows.

Twenty-four primiparous sows were used to determine the extent of mammary gland growth during lactation. Litter size was set to nine or 10 pigs immediately after birth. Sows were slaughtered in groups representing d 0 (within 12 h after farrowing), 5, 10, 14, 21, and 28 of lactation. Sows were provided 17.5 Mcal ME and 65 g of lysine per day during lactation. Mammary glands were collected at slaughter and trimmed of skin and extraneous fat pad. Each gland was weighed, cut in half to measure cross-sectional area, and ground for chemical analysis. Dry matter content, dry fat-free tissue (DFFT) content, protein content, amino acids composition, ash content, and DNA content were measured. Only glands known to have been suckled were included in these data. Wet and dry tissue weight; cross-sectional area; and the amount of DFFT, tissue protein, and amino acids in each suckled mammary gland increased (P < .05) during lactation to a peak on d 21. Fat percentage of each suckled gland declined (P < .05) and the percentage of protein and DFFT increased (P < .05) as lactation progressed. These results suggest that hypertrophy occurred in the tissue during lactation. There was a linear increase in the amount and percentage of DNA during lactation (P < .05), suggesting hyperplasia of the mammary tissue. Mammary tissue growth continues in suckled glands during lactation in sows, with gland wet weight increased by 55% and total gland DNA increased by 100% between d 5 and 21 of lactation.

Epidermal growth factor induces cell proliferation in mouse pancreas and salivary glands.

Epidermal growth factor (EGF) is a mitogenic stimulus in many tissues and occurs in large amounts in the pancreas and salivary glands. Whether EGF is mitogenic in the pancreas is controversial, and the EGF effect has not been studied in the salivary glands. Therefore, the aim of the present study was to investigate the possible effects on the pancreas and parotid and submandibular glands at different time intervals after exogenous EGF administration. Human recombinant EGF was infused subcutaneously by osmotic minipumps in three groups of mice (for 1, 3, and 7 days, respectively) at a dosage of 10 micrograms/kg/h (1.6 mumol/kg/h). Tritiated thymidine was infused intraperitoneally by osmotic minipumps for the same time periods, but only for the last 3 days in the 7-day group. After 1 day the pancreas increased in weight and the increase persisted throughout the study. No effect was seen on the parotid or submandibular gland wet weight. A slight transient increase in pancreas protein content was observed, whereas amylase content was unaffected. The labeling index of serous and ductal cells in the parotid gland increased from the third day. After 7 days, all cell types studied in the pancreas and parotid and submandibular glands were in a hyperproliferative state. The results show that EGF evoked a strong proliferative response on all cell types studied in the pancreas and parotid and submandibular glands.

Cell cycle control in isoproterenol-induced murine salivary acinar cell proliferation

The eukaryotic cell cycle is a summary of a complex network of signal transduction pathways resulting in both DNA replication and cell division. Cyclin-dependent kinases (CDKs) control the cell cycle in all eukaryotes, whereas other proteins, known as cyclins, act as their regulatory subunits. Chronic injection with isoproterenol (ISO) can induce acinar cell proliferation in rodent salivary glands. Cyclins and CDK proteins from control and ISO-treated murine parotid acinar cells were detected by using Western blotting techniques. By comparing the expression of these cell cycle regulatory kinases in the parotid acinar cell transition from a quiescent state to a hypertrophic state, we found rapid increases in the protein levels of all CDKs, cyclin D and proliferating cell nuclear antigen (PCNA). The highest protein levels for CDKs and cyclins appeared at about 72 hr of ISO stimulation and were coincident with the highest rate of increase in gland wet weight. After 72 hr, the increase of both cell cycle protein and gland wet weight began to subside. By using a co-immunoprecipitation method, the following cell cycle regulators (CDK-cyclin complexes) were detected, CDK4-cyclin D, CDK2-cyclin E, CDK2-cyclin A, and cdc2-cyclin B, along with an increase in kinase activity over control untreated animals. Additionally, we detected significant decreases in the newly isolated CDK inhibitor (CKI) p27 kip but not Wee 1 kinase. The increased levels of CKI correlated with a decrease in kinase activity of CDK/cyclin complexes by 144 hr of chronic isoproterenol treatment. Our data suggest that the holoenzymes for cell cycle control (cyclin-CDK complexes) function as a final regulatory mechanism leading to salivary gland acinar cell proliferation. The gradual decline in protein levels of the CDKs and cyclins after 3 days of chronic treatment further indicates that ISO-induced proliferation of parotid acinar cells is self-limiting and non-tumorigenic.

Physiological monitoring of contaminant effects in individual rock crabs, Hemigrapsus edwardsi: The ecotoxicological significance of variability in response

1) Heart rates and midgut gland wet weight dry weight ( WW DW ) ratios of rock crabs ( Hemigrapsus edwardsi) were measured in individuals collected along a contaminant gradient in Otago Harbour, New Zealand. 2) Median heart rates decreased by 15–20% while midgut gland WW DW ratios increased by 25–30% with increasing contamination of field sites. 3) Intra- and inter-individual variability in heart rate increased markedly at the most severely contaminated sites, as did inter-individual variability in midgut gland WW DW ratio. 4) The findings are discussed in the context of the use of the biomarker approach to address ecotoxicological problems.

Hormonal regulation of nuclear type II estrogen binding sites in the dorsolateral prostate of Noble rats

We previously demonstrated that simultaneous treatment of Noble (NBL) rats with estradiol (E2) and testosterone (T) for 16 weeks induces a proliferative response selectively in the dorsolateral prostates (DLP) of all treated animals [1, 2]. The unique sensitivity of rat DLP to the conjoint androgen-estrogen-induced mitogenic action may be attributable to the presence of a moderate affinity, high capacity, nuclear estrogen binder (type II sites) found exclusively in this prostatic lobe [2, 3]. Little is known about whether prostatic type II site levels are under hormonal regulation. The aim of this study is to determine whether testicular steroids play a role in regulating the basal and/or induced levels of type II site expression in rat DLP. In the first experiment, rats were castrated and immediately treated with 5 alpha-dihydrotestosterone (DHT) and/or E2 for 6 weeks to determine whether these steroids, separately or jointly, could sustain DLP type II site levels in castrates. Treatments of castrated rats with DHT and DHT+E2 were found to be effective in maintaining DLP type II site levels and gland wet weights at values close to those found in intact untreated controls, while treatments with E2 failed to maintain these parameters at levels observed in intact animals. In the second experiment, intact rats were treated with an androgen (T or DHT) or E2, alone or in combination, for 16 weeks to ascertain which hormonal regimen could induce a higher level of type II site expression in the DLP. Treatments of rats with an androgen (T or DHT) or E2 alone did not change DLP type II site levels even though T treatment caused a slight increase in gland weight, while E2 treatment induced prostatic atrophy. Contrary to single hormone treatments, combined T + E2 and DHT+E2 treatments were effective in inducing a doubling of type II sites and increases in wet weight of the DLPs. These data indicate that testicular androgen is the primary factor responsible for maintaining a basal level of type II site expression in rat DLP, while conjoint androgenic-estrogenic action is needed for the induction of a higher level of type II site expression in the tissue.

Modification of the β-adrenergic sensitivity of rat submandibular gland by constant light

Rats exposed for 2, 5, 10, 20, 35 and 50 days to constant light (CL) showed β-adrenergic desensitization of the submandibular gland as indicated by dose-response curves to 1.5, 2.0, 3.0, 10.0, 30.0 and 50.0 μg/kg isoproterenol. The phenomenon, evident with 1.5, 2.0 and 50.0 μg/kg after 2 days at CL, was more intense as time of exposure increased to reach values of about 50% those for controls after 20 days. After 35 and 50 days, desensitization showed some reversion, but the secretory responses were still lower than for controls maintained under a typical photoperiod. In the groups of rats exposed to CL for 20 days, gland wet and dry weights were higher (10%) than those of controls. These changes probably indicate an adaptation of the sympathetic control of salivary secretion induced by environmental illumination.

The effect of ecdysis on DNA of the hepatopancreas and green gland of the Florida spiny lobster ( Panulirus argus)

The DNA content per gram of wet hepatopancreas and green gland of Panulirus argus was found to vary with molt stage. In hepatopancreas of premolt and postmolt spiny lobsters, the DNA content was, respectively, 5-fold and 3-fold higher than in intermolt animals. Green gland DNA content showed a trend of varying in the same manner as hepatopancreas DNA. The DNA content per gram of wet green gland from intermolt lobsters averaged about 50 times higher than that of hepatopancreas. The changes in DNA content were not related to changes in wet tissue weights during the molting cycle. The ratio of hepatopancreas or green gland wet weights to total body weight remained constant. A ribosomal DNA probe hybridized to intermolt hepatopancreas DNA samples to a much greater extent than to premolt or postmolt samples, suggesting that rDNA sequences are under-represented in the amplified DNA which occurs prior to ecdysis.

Cholecystokinin modulates isoproterenol induced changes in rat parotid gland.

1. Parotid gland secretory function and activity of several enzymes involved in intracellular second messenger signalling were measured in rats receiving 0.5 ml i.p. injections of saline (control), isoproterenol, CCK or both drugs. 2. Isoproterenol caused a 2.5-fold increase in parotid gland wet weight compared to control. Chronic administration of CCK alone has no effect on gland weight. A combination of CCK and isoproterenol did not alter the hypertrophy of the gland observed with isoproterenol alone. 3. Isoproterenol administration caused a 74% decrease in parotid gland amylase enzyme activity. While CCK alone did not influence the enzyme activity, it depressed amylase mRNA steady state levels and had an additive effect on further decreasing mRNA levels when administered in combination with beta-agonist. 4. Phospholipase C registered an increase ranging from 22 to 38% in all experimental groups as compared to control. 5. Parotid gland protein kinase C and PdtIns 3-kinase activity were not altered in response to CCK alone, but in combination with isoproterenol, appeared to moderate beta-agonist signal transduction responses.

Effects of liquid diet on amylase activity and mRNA levels in rat parotid gland

Rats maintained on this diet demonstrated a dramatic reduction in amylase enzyme activity. This reduced level was evident through quantitation based on activity per mg gland protein, as well as on activity per gland wet weight. This was confirmed by the densitometric scanning of Western blots of sodium dodecyl sulphate-polyacrylamide gels of total parotid gland protein. Immune precipitation of enzyme activity from cell lysates showed that liquid-diet animals had lower levels of amylase than chow-fed controls. Isolation of poly (A +)-containing RNA followed by in vitro translation indicated that the steady-state level of amylase-specific mRNA had increased relative to the total mRNA pool. This outcome was also observed by dot-blot analysis of the cellular RNA using an amylase cDNA probe. Thus the regulation of amylase synthesis in rats maintained on liquid diets may take place at the level of protein translation, as well as of differential mRNA stability or gene transcription.

Morphological changes in the rabbit submandibular gland after parasympathetic or sympathetic denervation

Selective denervation was used to obtain further information about the relative roles of the nerves supplying the gland. Parasympathetic denervation soon caused atrophy in both acinar and granular tubule cells, and a substantial reduction in gland wet weight. The secretory cells had several unusual features, and striated duct cells tended to accumulate glycogen. Three to four weeks after pre-ganglionic parasympathectomy, the parenchymal cells were less abnormal but still smaller than in the control, unoperated gland. Thus, parasympathetic impulses are required to maintain normal parenchymal cells. Chronic sympathectomy caused little or no detectable changes in the secretory cells, and there was no significant change in wet weight. Nevertheless, pre-ganglionic sympathetic axotomy reduced the reflex secretion of acinar mucosubstances in response to feeding. Thus, sympathetic impulses normally contribute to the reflex secretion of acinar mucosubstance from these glands.

The effects of starvation and refeeding on egg laying and the synthetic activity of the albumen gland in Bulinus truncatus, a snail vector of urinary schistosomiasis

1. 1. The effects of starvation (for 1, 2, 3, 6, 9 and 12 days, respectively) and refeeding (12 days starvation and 1, 2 and 3 days refeeding, respectively) on egg laying and albumen gland activity in the freshwater snail, Bulinus truncatus were studied. 2. 2. The egg laying of starving snails rapidly decreased and ceased by day 6 of starvation. Egg laying was restored 24 hr after refeeding. The recorded decrease in albumen gland wet weight was proportional to the starvation periods. The DNA contents of the glands of the different experimental groups was not statistically different from the controls. 3. 3. Albumen gland synthetic activity expressed as 14C-glucose incorporation into galactogen/μg DNA and 3H-amino acids into total protein was determined. The glands showed an abrupt decrease in synthetic activity after 1 day of starvation and gradually decreased further until days 9–12. The decrease in activity of the glands was more rapid than that of egg laying. Upon refeeding, the activity of the glands recovered rapidly, simultaneous with the increase in wet weight and egg laying. 4. 4. In conclusion, there is a correlation between egg laying and the in vitro activity of albumen glands. The results show a short-term effect of starvation on the fecundity of the snails. Such studies could be useful in field studies as well as snail control by applying molluscicides under optimal conditions.

760 DIFFERENCES IN PAROTID SALIVA SODIUM LEVELS AND RESERPINE-RESPONSIVENESS BETWEEN RAT LINES

The mechanism underlying the inhibition of Na reabsorption from exocrine secretions in cystic fibrosis (CF) is not known. In the animal CF phenocopy of Martinez, chronic reserpine (RS) administration to rats causes histological and physiological changes in exocrine glands similar to those in CF. To examine the effect of chemical sympathectomy on Na transport, we studied the onset of inhibition of Na reaborption from parotid saliva in male Sprague-Dawley (SD) rats given i.p. RS (0.5 mg/kg) daily. Saliva stimulated by i.p. pilocarpine was collected by cannulating the parotid duct. A single RS dose caused a marked increase in Na content within 24 hr in rats from our colony (OU). Na content increased linearly with the duration of RS administration. Saliva of SD rats from Sasco Inc. (SI) contained much less Na than that of OU rats at all saliva flow rates below 70 ul/min/g gland wet weight. RS administered to SI rats caused no demonstrable increase in saliva Na content until the 4th day. Na increased only to levels seen in untreated OU rats. These results indicate that both the intrinsic activity and the susceptibility to inhibition of the ductal Na transport system of exocrine glands show specific differences. Such differences complicate the use of uncharacterized rat strains for the bioassay of CF factors but may provide valuable insights into the biological regulation of Na transport in the exocrine gland duct.

Effect of a ground versus a pelleted bulk diet on the rat parotid gland

Rats were fed a stock pelleted diet (control diet) or the same stock diet to which 30 per cent non-nutritive cellulose had been added (bulk diet). The control diet and the bulk diet were fed to rats in two consistencies, pelleted or ground. Compared to the parotid glands of rats fed the pelleted control diet, those of rats fed either of the ground diets were not different, but parotid gland wet weight, DNA and RNA content and total gland amylase and protein were significantly greater in rats fed the pelleted bulk diet for 2 wk. The cell indices of RNA/ DNA and amylase/DNA were significantly greater in rats fed the pelleted bulk diet, but were not significantly changed in rats fed the ground bulk diet. Thus there is a significant increase in cell number and cell content of amylase when rats are fed a bulk diet that is pelleted and no such increases are found when the bulk diet is ground. The findings support the contention that the physical consistency of the diet and its requirement for mastication are major factors contributing to the regulation of parotid-gland size and secretory-enzyme content.

Effects of the Sex Steroid Hormones and Vitamin D3 on Calcium-Binding Proteins in the Chick Shell Gland1

The effects of diethylstilbestrol (DES), progesterone, testosterone and vitamin 03, alone or in various combinations, on the calcium-binding protein (CaBP) concentration, wet weight and histology of the shell gland were investigated in 7.5-10-week-old chicks. Chicks, which were either vitamin D deficient or vitamin D replete, were pretreated with DES to differentiate the shell gland. DES administration was then discontinued for 10 days and then chicks were treated with the hormones to be tested. In this secondary treatment period, DES increased the levels of CaBP (measured using a specific radioimmunodiffusion assay) in both vitamin D deficient and vitamin D replete chicks, suggesting an effect of DES on CaBP largely independent of vitamin 0. Progesterone alone had no effect on CaBP levels, but it inhibited the DES stimulated increase in CaBP in every case tested. In contrast to this antagonism of CaBP level, the increases in shell gland wet weight and hypertrophy of the tubular gland cells induced by DES were not inhibited by concomitant progesterone administration. Vitamin D3 alone did not increase CaBP concentration in the shell gland and, when administered with DES to vitamin D deficient chicks, it elicited little if any increase in CaBP level above that evoked by DES alone. Testosterone had little effect on CaBP level in the shell gland. Despite chronic injections with estrogen, however, the maximum concentration of CaBP induced in the DES-differentiated shell gland was only one-sixth the concentration found in the shell gland of the egg laying hen. These results indicate that CaBP, which is found in numerous organs, may be regulated by factors in addition to vitamin D3, the predominant regulator in the intestine, and that estrogen and progesterone may play important roles in modulation of CaBP concentration in the shell gland of the hen.

Effects of prolactin and progesterone on preputial gland growth and progesterone metabolism in hypophysectomized, ovariectomized immature rats

Using a 2 × 2 factorial experimental design, treatments of hypophysectomized, ovariectomized, immature rats with prolactin (100 μg twice daily) and progesterone (2mg/day) resulted in a significant synergism between the two hormones on preputial gland wet weights. Prolactin or progesterone treatment alone, caused gland wet weights to be significantly greater than those of controls. Investigations of the possible modes of action of prolactin on the preputial gland showed that prolactin decreased serum testosterone and/or 17β-hydroxy-5α-androstan-3-one levels but had no effect on serum progesterone levels, indicating that the effect of prolactin is not mediated via an elevation in these serum steroid levels. Prolactin increased the overall rate of metabolism of [ 14C]-progesterone by preputial gland tissue in vitro in proportion to its ability to increase wet weights of the glands, but did not alter the pattern of this metabolism. In vivo progesterone treatment caused a significant change in the pattern of [ 14C]-progesterone metabolites in vitro. The main metabolites whose rates of formation were increased by progesterone treatment, were identified as 20α-hydroxy-5α-pregnan-3-one and 5α-pregnan-3α-20α-diol along with its 3β epimer. A less abundant metabolite, whose formation was diminished by progesterone treatment, was identified as 3α-hydroxy-5α-pregnan-20-one. The removal of the adrenal glands did not alter the effects of in vivo progesterone treatment on the preputial gland. Therefore, the data indicate that prolactin and progesterone exert separate effects on the preputial gland of the immature rat.