Physiology of ghrelin and related peptides.

Growth hormone (GH) released from pituitary under direct control of hypothalamic releasing (i.e., GHRH) and inhibiting (i.e., sst or SRIF) hormones is an anabolic hormone that regulates metabolism of proteins, fats, sugars and minerals in mammals. Cyril Bowers' discovery of GH-releasing peptide (GHRP-6) was followed by a search for synthetic peptide and nonpeptide GH-secretagogues (GHSs) that stimulate GH release, as well as a receptor(s) unique from GHRH receptor. GHRH and GHSs operate through distinct G protein-coupled receptors to release GH. Signal transduction pathways activated by GHS increase intracellular Ca2+ concentration in somatotrophs, whereas GHRH increases cAMP. Isolation and characterization of ghrelin, the natural ligand for GHS receptor, has opened a new era of understanding to physiology of anabolism, feeding behavior, and nutritional homeostasis for GH secretion and gastrointestinal motility through gut-brain interactions. Other peptide hormones (i.e., motilin, TRH, PACAP, GnRH, leptin, FMRF amide, galanin, NPY, NPW) from gut, brain and other tissues also play a role in modulating GH secretion in livestock and lower vertebrate species. Physiological processes, such as neurotransmission, and secretion of hormones or enzymes, require fusion of secretory vesicles at the cell plasma membrane and expulsion of vesicular contents. This process for GH release from porcine somatotrophs was revealed by atomic force microscopy (AFM), transmission electron microscopy (TEM) and immunohistochemical distribution of the cells in pituitary during stages of development.

Interaction of talin with actin: sensitive modulation of filament crosslinking activity.

Talin is an adhesion plaque protein believed important in linking actin filaments to the plasma membrane. The nature of a direct talin-actin interaction, however, is complex and has remained unclear. We have systematically characterized the effects of pH, ionic strength, temperature, and protein molar ratio on the interaction between highly purified talin and actin. The ability of talin to increase viscosity of F-actin at 25 degrees C and low ionic strength increased with decreasing pH from 7.3 to 6.4 and increasing molar ratio of talin to actin. At pH 6.4 and low ionic strength, talin could extensively crosslink actin filaments into ordered bundles as shown by negative staining and could cosediment with F-actin at molar ratios as high as one talin to two actin monomers. Talin crosslinked prepolymerized actin filaments to a similar extent as actin filaments polymerized in its presence. The 190-kDa calpain-generated proteolytic fragment of talin bound poorly to actin under conditions favorable for intact talin, but was able to crosslink actin filaments at a lower pH. Increasing the ionic strength within a relatively narrow range significantly decreased ability of talin to bind to actin, regardless of pH. The effects of pH and ionic strength on the talin-actin interaction were rapid and reversible. Low-shear-viscosity studies revealed a strong temperature dependence in the talin-actin interaction with significant crosslinking activity at physiological-like ionic conditions and temperature (37 degrees C). Our results consistently demonstrated that talin crosslinks actin filaments and that this direct interaction is highly sensitive to, and dependent upon, ionic conditions and temperature.

Properties of the novel intermediate filament protein synemin and its identification in mammalian muscle.

We examined specific properties of highly purified synemin (230 kDa), recently identified as a novel intermediate filament (IF) protein, from avian smooth muscle. Soluble synemin in 10 mM Tris-HCl, pH 8.5, appears as approximately 11-nm-diameter globular structures by negative-stain and low-angle shadow electron microscopy. Chemical crosslinking and SDS-PAGE analysis indicate that soluble synemin molecules contain two 230-kDa subunits. The pH- and ionic strength-dependent solubility properties of synemin are similar to those of the type III IF protein desmin, but under physiological-like conditions in which desmin self-assembles into long approximately 10-nm-diameter IFs, synemin self-associates into complex, approx 15- to 25-nm-diameter globular structures. Calpain digestion demonstrated that synemin is extremely proteolytically labile. Western blot analysis, with monospecific polyclonal antibodies against avian synemin, shows the presence of the reactive 230-kDa synemin band in samples of adult avian skeletal, cardiac, and smooth muscle and of two reactive bands at approximately 225 kDa (major) and approximately 195 kDa in adult porcine skeletal, cardiac, and smooth muscle. Partial purification of synemin from porcine smooth muscle also resulted in fractions highly enriched in the approximately 225- and approximately 195-kDa polypeptides. Conventional immunofluorescence and immunoconfocal microscopy of isolated myofibrils and of frozen sections also demonstrated, for the first time, that synemin is present in all three adult porcine muscle cell types and is colocalized with desmin in skeletal and cardiac muscle cells at the myofibrillar Z-lines.

The cytoskeleton in skeletal, cardiac and smooth muscle cells.

The muscle cell cytoskeleton consists of proteins or structures whose primary function is to link, anchor or tether structural components inside the cell. Two important attributes of the cytoskeleton are strength of the various attachments and flexibility to accommodate the changes in cell geometry that occur during contraction. In striated muscle cells, extramyofibrillar and intramyofibrillar domains of the cytoskeleton have been identified. Evidence of the extramyofibrillar cytoskeleton is seen at the cytoplasmic face of the sarcolemma in striated muscle where vinculin- and dystrophin-rich costameres adjacent to sarcomeric Z lines anchor intermediate filaments that span from peripheral myofibrils to the sarcolemma. Intermediate filaments also link Z lines of adjacent myofibrils and may, in some muscles, link successive Z lines within a myofibril at the surface of the myofibril. The intramyofibrillar cytoskeletal domain includes elastic titin filaments from adjacent sarcomeres that are anchored in the Z line and continue through the M line at the center of the sarcomere; inelastic nebulin filaments also anchored in the Z line and co-extensible with thin filaments; the Z line, which also anchors thin filaments from adjacent sarcomeres; and the M line, which forms bridges between the centers of adjacent thick filaments. In smooth muscle, the cytoskeleton includes adherens junctions at the cytoplasmic face of the sarcolemma, which anchor beta-actin filaments and intermediate filaments of the cytoskeleton, and dense bodies in the cytoplasm, which also anchor actin filaments and intermediate filaments and which may be the interface between cytoskeletal and contractile elements.

Integrin expression in developing smooth muscle cells.

We studied the specific expression patterns and distributions of alpha1 and beta1 integrin subunits, the major cell adhesion receptors in smooth muscle, in developing smooth muscle cells from 16-, 18-, and 20-day embryonic gizzards and from 1- and 7-day post hatch chick gizzards by SDS-PAGE, immunoblotting, and immunoelectron microscopy. Antibodies raised against alpha1 and beta1 integrins isolated from avian gizzards were used as probes. Gels and blots showed that the amount of alpha1 and beta1 integrins increased as age increased, with major increases at 1 and 7 days post hatch. Image analysis of immunoelectron micrographs demonstrated that statistically significant labeling increases occurred between embryonic Days 16 and 18, between embryonic Day 20 and 1 day post hatch, and between 1 day and 7 days post hatch. Immunolabeling with both anti-alpha1 and anti-beta1 integrin was prominent at membrane-associated dense plaques (MADPs) and at filament anchoring regions at cell ends. This indicates that alpha1 and beta1 integrin expression coincides temporally with the intracellular proliferation and reorientation of myofilaments. The similarity in distribution patterns of alpha1 and beta1 integrins during development suggests that the two integrin subunits are synchronously expressed during development and do not appear sequentially. (J Histochem Cytochem 46:119-125, 1998)

Effects of electrical stimulation and postmortem storage on changes in titin, nebulin, desmin, troponin-T, and muscle ultrastructure in Bos indicus crossbred cattle.

The effects of electrical stimulation (ES) on degradation of titin, nebulin, desmin, and troponin-T (TN-T) and on structural changes in the longissimus muscle (LM) from Brahman x Simmental (B x S) cattle (Bos indicus cross) were determined. The left side of seven B x S beef carcasses was stimulated (200 V, 20 Hz) within 1 h of death, and the right side was the nonstimulated (NS) control. Myofibrils for SDS-PAGE and samples for transmission electron microscopy were prepared from the LM at 0, 1, 3, 7, 14, and 28 d postmortem (PM). The SDS-PAGE results showed that the T1 band of titin was absent by 7 d in two animals, by 14 d in four animals, and by 28 d in one animal in both NS and ES samples. By SDS-PAGE, intact nebulin was gone by 7 d in two animals and by 14 d in five animals, but in blots, nebulin decreased by 7 d and was absent by 14 d in both NS and ES samples. The desmin band could still be seen as a light band at 28 d in Western blots of both NS and ES samples. A decrease in TN-T and a concomitant increase in the 30-kDa polypeptide were observed in both NS and ES samples. Western blots with a monoclonal antibody to TN-T confirmed that TN-T decreased at similar rates in NS and ES samples but showed that the 30-kDa polypeptide was more heavily labeled in ES samples from 7 to 28 d. Contraction nodes were present in O-d ES samples and were still observed in 28-d ES samples. Narrow, intermediate, and wide I-band fractures were seen earlier and at a greater frequency in ES than in NS samples. Overall, ES had no detectable effect on titin, nebulin, desmin, or TN-T degradation but accelerated the appearance and enhanced the frequency of three types of I-band fractures in the LM from Bos indicus crossbred cattle.

Effect of electrical stimulation on postmortem titin, nebulin, desmin, and troponin-T degradation and ultrastructural changes in bovine longissimus muscle.

Electrical stimulation (ES) of bovine carcasses is usually done to increase tenderness and has been hypothesized to increase the activity of proteolytic enzymes that may degrade structural proteins in muscle cells and cause fractures and breaks in muscle fibers, thus enhancing meat tenderness. Our objective was to compare postmortem (PM) changes in the muscle proteins, titin, nebulin, alpha-actinin, desmin, and troponin-T and in myofibrillar structure in nonstimulated (NS) and ES bovine skeletal muscle. One side of eight beef carcasses was stimulated within 1 h of death, and the other side was the NS control. Myofibrils for SDS-PAGE and samples for transmission electron microscopy were prepared from the longissimus muscle at 0, 1, 3, 7, 14, and 28 d PM. In SDS-PAGE, titin migrated as three bands in both NS and ES 0-d samples. The slowest migrating band, T1 (intact titin), decreased slightly faster in ES samples from five animals. The fastest migrating band, T2 (degraded titin), increased in amount through 3 d and was still present at 28 d. A titin monoclonal antibody (mAb) identified a large family of degradation products that migrated faster than myosin heavy chains and that was more heavily labeled in Western blots of ES samples than in NS samples. In SDS-PAGE of NS samples, intact nebulin disappeared by 3 d in three animals, by 7 d in four animals, and by 14 d in one animal, but in ES samples the nebulin band was absent by 3 d in three animals and by 7 d in five animals. SDS-PAGE showed that the amount of intact desmin decreased slightly sooner in two ES samples and was absent earlier in one ES sample than in the corresponding NS control samples. Blots labeled with a polyclonal antibody to desmin showed that a more heavily labeled 38-kDa desmin degradation product was present in ES than in NS samples. Postmortem degradation of alpha-actinin was not detected. Contraction node (CN) formation, stretching of conjoined sarcomeres adjacent to the nodes, increased frequency of I-band fractures and accelerated appearance of wide I-band fractures adjacent to the Z-line, and, in some animals, slightly accelerated degradation of titin, nebulin, and troponin-T were characteristics of ES muscle.

Talin can crosslink actin filaments into both networks and bundles.

The talin-actin interaction was examined by using negative staining and cosedimentation assays. At pH 6.4 and low ionic strength, talin extensively crosslinked actin filaments into both networks and bundles. The bundles consist of parallel actin filaments with a center-to-center distance of 13 nm, and talin crossbridges spaced at 36-nm intervals along the bundles. As pH was increased stepwise from 6.4 to 7.3, talin's bundling activity was decreased first, then its networking activity. Qualitatively similar results were obtained at pH 6.4 by increasing ionic strength. Chemical crosslinking indicated talin was present as a dimer from pH 6.4 to 7.3, with or without added KC1. The results show that talin can interact directly with actin filaments by formation of actin filament networks and bundles, with the bundles more sensitive to dissolution by increase in pH or ionic strength.

Immunocytochemistry of the muscle cell cytoskeleton.

The muscle cell cytoskeleton is defined for this review as any structure or protein primarily involved in linking or connecting protein filaments to each other or to anchoring sites. In striated muscle, the M line connects thick filaments at their centers to adjacent thick filaments. Titin forms elastic filaments that extend from the M line to the Z line and may contribute to the resting tension properties of striated muscle. Nebulin forms inextensible filaments in skeletal muscle that are closely associated with thin filaments and that may provide a length template for thin filaments. Z lines anchor thin filaments from adjacent sarcomeres via the actin-binding function of alpha-actinin. Other proteins located at the Z line include Cap Z, Z-nin, Z protein, and zeugmatin. Intermediate filaments connect myofibrils to each other at the level of the Z line and to the sarcolemma at the Z- and possibly the M-line levels. Immunolocalization has identified the adhesion plaque proteins spectrin, vinculin, dystrophin, ankyrin, and talin at subsarcolemmal sites where they may be involved with filament attachment. Smooth muscle cell cytoskeletons are believed to include membrane associated dense bodies (MADBs), intermediate filaments, cytoplasmic dense bodies (CDBs), and perhaps a subset of actin filaments. MADBs contain a menu of attachment plaque proteins and anchor both thin filaments and intermediate filaments to the sarcolemma. CDBs are intracellular analogs of striated muscle Z lines and anchor thin filaments and intermediate filaments.

Identification of the 30 kDa polypeptide in post mortem skeletal muscle as a degradation product of troponin-T.

Although a 30 kDa polypeptide frequently is seen by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of post mortem (pm) skeletal muscle and in turn is used as an indicator of proteolysis, its origin has not been conclusively identified. We used antibodies to selected myofibrillar proteins, including some known to be degraded pm, to identify this polypeptide. The left side of eight beef carcasses was electrically stimulated (ES) within 1 h after slaughter, and the right side served as the non-stimulated (NS) control. The longissimus lumborum (LL) muscle was removed from the carcass at 24 h pm and was stored at 2 degrees C. Myofibrils were prepared from the LL muscle immediately after stimulation (0 day) and from the stored muscle sample at 1, 3, 7, 14 and 28 days pm for analysis of SDS-PAGE and Western blots. By SDS-PAGE, troponin-T (TN-T) decreased in amount more rapidly pm in ES samples than in NS samples. By SDS-PAGE, a 30 kDa band increased and became a prominent band by 7 days pm in both NS and ES samples. A monoclonal antibody (mAb) to TN-T labeled purified TN-T, as well as the TN-T in myofibrils, a prominent 30 kDa polypeptide and a family of lower molecular mass polypeptides in pm muscle. This mAb also labeled a 30 kDa band that had been electrophoretically purified from pm muscle.(ABSTRACT TRUNCATED AT 250 WORDS)

Substructure of cytoplasmic dense bodies and changes in distribution of desmin and alpha-actinin in developing smooth muscle cells.

The substructure of assembling cytoplasmic dense bodies (CDBs) and changes in the distribution of desmin and alpha-actinin during development of smooth muscle were studied in gizzard samples from 10- and 16-day embryos and from 1- and 7-day post-hatch chickens. CDBs in these cells lack the density of CDBs in mature or adult smooth muscle cells and, thus, allow observations of the changes inside CDBs. The random filament orientation seen in younger embryonic cells is first modified to include relatively small patches of IFs that are somewhat straighter and are approaching a side-by-side arrangement. As development proceeds, the IFs in these arrays become straighter, are parallel over longer lengths of the IFs and later acquire the density characteristic of mature CDBs. Anti-desmin labeling in embryonic 10- and 16-day cells showed that desmin intermediate filaments (IFs) were located in the myofilament compartment but were concentrated in or near assembling CDBs. Anti-desmin labeling shifted to the perimeter of CDBs after hatching. Cross sections, longitudinal sections, and stereo pairs all show that IF profiles are present inside unlabeled assembling CDBs. Anti-alpha-actinin labeling was directly on CDBs and was often associated with the cross-connecting filaments (CCFs) (average diameter of 2-3nm) inside CDBs. We propose, based on these data, that desmin IFs, alpha-actinin-containing CCFs, and actin filaments are the principal components of the substructure of assembling CDBs. We also present a proposed model for CDB assembly.

The marked pH dependence of the talin-actin interaction.

The effect of pH on the interaction between talin and actin was examined by using cosedimentation assays, low shear viscometry, and electron microscopy. When the pH at which the interaction was tested was lowered from 7.5 to 7.0 and below, the ability of talin to cosediment with F-actin was greatly enhanced, with maximal binding at pH 6.6. Talin's ability to increase the low shear viscosity of F-actin solutions was also significantly elevated as pH was decreased from 7.4 to 6.6. Electron microscope observations of negatively stained actin controls and talin-actin mixtures supported these results by demonstrating an increase in the degree of crosslinking of actin filaments in the presence of talin at pH values of 7 and lower.

Synergistic effects of insulin-like growth factor I and gonadotrophins on relaxin and progesterone secretion by ageing corpora lutea of pigs.

Insulin-like growth factor I (IGF-I) is involved in paracrine/autocrine regulation of gonadal steroidogenesis and peptide hormone biosynthesis. This study was designed to determine whether IGF-I alone, or an interaction of IGF-I, is involved in augmenting the actions of luteinizing hormone (LH) and prolactin in controlling relaxin and progesterone secretion from ageing corpora lutea of hysterectomized gilts at days 110, 113 and 116 after oestrus. Luteal tissue slices were incubated for 8 h with IGF-I (0, 50, 300 ng ml-1), LH (0, 100, 1000 ng ml-1), and prolactin (0, 100, 1000 ng ml-1) alone or in combination. Progesterone and relaxin concentrations were determined by radioimmunoassay of spent medium and of homogenates from luteal tissue slices before and after incubation. Porcine luteal tissue from day 110 had a net output of 25 ng progesterone and 26 ng relaxin in the control and of 65 ng progesterone and 2125 ng relaxin in the combined IGF-I, LH and prolactin treatment mg-1 of luteal tissue, respectively. IGF-I, LH and prolactin alone or in combination significantly increased (P < 0.01) progesterone production by luteal tissue from day 110, but they were partially effective at day 113 and ineffective at day 116. By contrast, the same hormone treatments increased relaxin production by luteal tissue from days 110 and 113. Even at day 116, prolactin alone or with LH or IGF-I continued to stimulate relaxin production. In conclusion, IGF-I augments the ability of prolactin and LH to increase relaxin production by ageing corpora lutea; however, a decrease in progesterone secretion and an increase in relaxin secretion at day 113 indicate that different mechanisms control progesterone and relaxin secretion in pigs.

Assembly of contractile and cytoskeletal elements in developing smooth muscle cells.

Specific developmental changes in smooth muscle were studied in gizzards obtained from 6-, 8-, 10-, 12-, 14-, 16-, 18-, and 20-day chick embryos and from 1- and 7-day posthatch chicks. Myoblasts were actively replicating in tissue from 6-day embryos. Cytoplasmic dense bodies (CDBs) first appeared at Embryonic Day 8 (E8) and were recognized as patches of increased electron density that consisted of actin filaments (AFs), intermediate filaments (IFs), and cross-connecting filaments (CCFs). Although the assembly of CDBs was not synchronized within a cell, the number, size, and electron density of CDBs increased as age increased. Membrane-associated dense bodies (MADBs) also could be recognized at E8. The number and size of MADBs increased as age increased, especially after E16. Filaments with the diameter of thick filaments first appeared at E12. Smooth muscle cells were able to divide as late as E20. The axial intermediate filament bundle (IFB) could first be identified in 1-day posthatch cells and became larger and more prominent in 7-day posthatch cells. Immunogold labeling of 1- and 7-day posthatch cells with anti-desmin showed that the IFB contained desmin IFs. The developmental events during this 23-day period were classified into seven stages, based primarily on the appearance and the growth of contractile and cytoskeletal elements. These stages are myoblast proliferation, dense body appearance, thick filament appearance, dense body growth, muscle cell replication, IFB appearance, and appearance of adult type cells. Smooth muscle cells in each stage express similar developmental characteristics. The mechanism of assembly of myofilaments and cytoskeletal elements in smooth muscle in vivo indicates that myofilaments (AFs and thick filaments) and filament attachment sites (CDBs and MADBs) are assembled before the axial IFB, a major cytoskeletal element.

Abrupt shifts in relaxin and progesterone secretion by aging luteal cells: luteotropic response in hysterectomized and pregnant pigs.

Although a prominent role of conceptuses in the hormonal control of normal parturition in the pig is well founded, mechanisms are entrained that cause a timed relaxin release and an abrupt decrease in progesterone secretion in the complete absence of the uterus. This study focused on the steroidogenic and peptide hormone secretory capacities of aging porcine luteal cells in culture on specific days during a narrow window (days 110-116) when abrupt shifts occur in their function. Dispersed luteal cells (3 x 10(5) cells/well) from hysterectomized and pregnant gilts on days 110, 113, and 116 after estrus were cultured for 24 h with porcine (p) LH, pPRL, prostaglandin E1 (PGE1), PGE2, and (Bu)2cAMP. Progesterone and relaxin concentrations in cells preceding culture and in the spent medium were quantified by RIA. Progesterone in cells before culture was less (P less than 0.05) than that released into the medium, while relaxin levels were greater (P less than 0.05) than those in the medium. Relaxin stored in luteal cells of hysterectomized gilts was consistently greater (P less than 0.01) than that during the same days of late pregnancy; however, progesterone was greater (P less than 0.01) in hysterectomized compared with pregnant gilts only on day 116. Relaxin release was greater (P less than 0.01) in hysterectomized compared with pregnant gilts on days 113 and 116, whereas progesterone was greater (P less than 0.05) on days 110 and 116 in hysterectomized compared with pregnant animals. pLH, pPRL, and (Bu)2cAMP increased (P less than 0.05) relaxin secretion in hysterectomized gilts on days 110, 113, and 116, but only on day 110 in pregnant animals. These hormones consistently increased (P less than 0.05) progesterone release only on day 110 in both groups of gilts. PGE1 increased (P less than 0.01) relaxin release in hysterectomized, but not pregnant, gilts; it was not effective in altering progesterone in either group. PGE2 increased the release of both hormones on day 113 in both groups. The percentage of large cells (i.e. greater than 20 microns in diameter) was consistently greater (P less than 0.01) on days 110-116 in hysterectomized compared with pregnant gilts. Both luteal cell relaxin concentrations and the population of electron-dense granules decreased from days 110-116 in pregnant, but not in hysterectomized, gilts; luteal cells from postpartum animals were nearly depleted of granules, and empty vesicles were prominent. The secretory response of the cells to tropic hormone stimulation is preprogrammed from the previous estrus that occurred more than 110 days previously.(ABSTRACT TRUNCATED AT 400 WORDS)

Determination of the critical concentration required for desmin assembly.

The critical concentration required for filament assembly in vitro from highly purified desmin was determined by both turbidity and centrifugation assays. Assembly was done in the presence of 2 mM-Ca2+, 2 mM-Mg2+ or 150 mM-Na+ at 2, 22 and 37 degrees C. Similar values for critical concentration were obtained by both assays. As temperature increased, critical concentration decreased for each cation. The critical concentration was lowest in the presence of Ca2+ at 2, 22 and 37 degrees C, but was highest in the presence of 150 mM-Na+ at 2 degrees C. Negative staining showed that supernatants from the centrifugation assays contained protofilaments, protofibrils and short particles (less than 300 nm), but pellets contained long filaments (greater than 1 micron) with an average diameter of 10 nm. As the temperature increased, both the average diameter and average length of particles in the supernatant increased. Thermodynamic analysis indicated that hydrophobic interactions were dominant during desmin assembly, but that ionic interactions might also be involved. Our results demonstrated that the specific cation and temperature and temperature-cation interactions all are important in assembly of desmin intermediate filaments.

Immunocytochemical identification of cytoskeletal linkages to smooth muscle cell nuclei and mitochondria.

In avian smooth muscle cells, desmin-containing intermediate filaments (IFs) are a prominent component of the cytoskeleton and are readily seen in several domains, including the axial intermediate filament bundle (IFB). Both the nucleus and some of the mitochondria are partly surrounded by elements of the IFB. By using anti-desmin and protein-A-colloidal gold labeling, we have identified intermediate filaments that form linkages with the nuclear envelope and with mitochondria. These linkage regions seem to occupy a proportionately greater part of the mitochondrial surface than of the nuclear envelope. The existence of these linkages in smooth muscle cells is consistent with results that support similar linkages to mitochondria and other cellular structures in various cells that contain either vimentin or keratin IFs. These linkages could functionally restrain or assist in homeostatically restoring organelles to their normal position after the rearrangement that accompanies the substantial shortening of smooth muscle cells.

Progesterone secretion and mitochondrial size of aging porcine corpora lutea.

A functional dependency between the nongravid uterus and the ovaries is essential to luteolysis and the return to estrus in the pig. After mating of gilts, the corpora lutea develop, and they are required for the maintenance of pregnancy to a normal duration of about 114 days. Hysterectomy of luteal phase (day 6) nongravid gilts results in persistence of the corpora lutea to 150 days. We report that these corpora secrete greater quantities (P less than 0.025) of progesterone than during the later half of gestation (days 54-108). Although aging corpora lutea remain functional for at least an additional 35 days, an abrupt reduction by half in progesterone secretion (16 ng/ml) occurs about day 114 in hysterectomized gilts that coincides with the prepartum decrease to basal serum levels (less than 0.5 ng/ml) at parturition (day 114) and during lactation. Aging corpora lutea remain large (averaging greater than 450 mg) on days 124 and 136 in hysterectomized gilts, whereas they regress (averaging less than 75 mg) in the lactating dams. Mitochondria continue to increase in size in aging corpora lutea of hysterectomized gilts until day 136; in contrast, they decrease during the postpartum period in lactating dams. A precisely timed signal, possibly of ovarian origin or from the CNS and pituitary gland, entrains in hysterectomized and pregnant pigs at day 113 that results in marked shifts in relaxin and progesterone secretion. Progesterone secretion and mitochondrial features suggest that porcine corpora lutea seem genetically controlled and are preprogrammed at estrus for the duration of pregnancy, regardless of the presence of conceptuses or absence of the uterus.