Regulation of membrane trafficking and endocytosis by protein kinase C: emerging role of the pericentrion, a novel protein kinase C-dependent subset of recycling endosomes.

The protein kinase C (PKC) family of isoenzymes has been shown to regulate a variety of cellular processes, including receptor desensitization and internalization, and this has sparked interest in further delineation of the roles of specific isoforms of PKC in membrane trafficking and endocytosis. Recent studies have identified a novel translocation of PKC to a juxtanuclear compartment, the pericentrion, which is distinct from the Golgi complex but epicentered on the centrosome. Sustained activation of PKC (longer than 30 min) also results in sequestration of plasma membrane lipids and proteins to the same compartment, demonstrating a global effect on endocytic trafficking. This review summarizes these studies, particularly focusing on the characterization of the pericentrion as a distinct PKC-dependent subset of recycling endosomes. We also discuss emerging insights into a role for PKC as a central hub in regulating vesicular transport pathways throughout the cell, with implications for a wide range of pathobiologic processes, e.g. diabetes and abnormal neurotransmission or receptor desensitization.

Linear Percoll gradient centrifugation of rat anterior pituitary cells. A simple method for prolactin cell enrichment.

Prolactin (PRL) cells were purified from nulliparous normal female adult Wistar rat pituitary cell suspensions by linear Percoll density gradient centrifugation, a procedure yielding single cells. Lactotrophs were found in two different layers, the first containing 70% PRL cells in the density range 1.055 to 1.065 g/ml, the second with 28% PRL cells in the range 1.070 to 1.080 g/ml. Both cell fractions contained more than 90% viable cells with an intact ultrastructure. The physiological integrity of the 70% enriched PRL cells was assessed by their basal PRL secretion, their secretory response to TRH and dopamine, and their cAMP production in a basal situation and after incubation with dopamine.

In vivo and in vitro effect of naloxone on prolactin response to TRH in rat.

In adult male Wistar rats submitted to a standardized noise stress, intravenous TRH induced a prolactin (PRL) secretory response. Prior IV naloxone administration not only lowered plasma PRL levels in those stressed rats but abolished also the stimulatory action of TRH. This effect was further studied by superfusion experiments on enriched PRL cell suspensions (70% lactotrophs) from female adult Wistar rats. Naloxone kept unaffected the basal PRL secretion but lowered significantly that induced by TRH. These experiments suggest a dual effect of naloxone on rat PRL secretion, one exerted on central opioid receptors lowering stress-related increased basal PRL levels, the other inhibiting the TRH-dependent PRL secretion exerted at the lactotroph level itself.

Postnatal development of growth hormone and prolactin cells in male and female rat pituitary. An immunocytochemical light and electron microscopic study.

Localization and ultrastructural maturation of prolactin (PRL) and growth hormone (GH) cells were studied in pituitaries from neonatal, immature (4-6 weeks old), and adult rats (2-3 months old) by light and electron microscopic immunocytochemistry. The distribution pattern of these cells did not change with age. Both cell types were concentrated laterodorsally, with PRL cells adjacent to the intermediate lobe and GH cells nearer the center of the pars distalis. Labeling density of the immunogold reaction was highest for both hormones in immature rats. In neonatal and immature rats, one PRL cell type with granules 200 nm in diameter was present. In adult rats, two types of PRL cells were present: one containing polymorphous granules measuring about 500 nm (prevalent in female rats), the other with spherical granules about 200 nm (prevalent in male rats). No changes were detected in GH cells during maturation.

Immunoreactive calcitonin content in foetal thyroid glands and in placentae of rats.

Immunoreactive calcitonin (I-CT) was assayed in rat thyroid glands from foetuses with a gestational age of 15.5-22 days. The mean content of the hormone was 455.0 +/- 35.8 ng/gland. A positive linear correlation was found between the I-CT content in the thyroid glands and the gestational age of the foetuses. I-CT was also present in rat placentae from 10.5-21.5 days old pregnancies. A mean placental concentration of 4.5 +/- 1.2 ng I-CT/mg wet tissue was found. The highest hormone concentrations were observed in placentae from 10.5 day old foetuses, displaying a progressive fall during the further course of pregnancy. It is concluded that the rat foetal thyroid gland produces I-CT beginning from a gestational age of 15.5 days'. The need for further investigations into a possible role of CT in transplacental calcium transport is also indicated.

Thyrotropin stimulated colloid droplet formation in the thyroid of hypophysectomised mice after calcitonin administration.

The influence of calcitonin on colloid droplet formation in the thyroid gland of hypophysectomised mice after thyrotropin (TSH) stimulation was investigated. Calcitonin was injected 30 min before TSH administration. 30 min after TSH the colloid droplet index in the thyroid cells was twice as low in the calcitonin group in comparison with the control one (0.111 +/- 0.020, 0.228 +/- 0.025 respectively). 60 min after thyrotropin treatment the number of colloid droplets was higher in the animals treated with calcitonin (0.135 +/- 0.093) in comparison with the control group (0.083 +/- 0.025). The possible mechanism of calcitonin action on colloid droplet formation is discussed.

Changes in serum TSH level after intraventricular injection of various neuromediators in rats.

The influence of various neuromediators on pituitary TSH secretion in rats has been investigated. Noradrenaline 50 microgram/rat, dopamine 50 microgram/rat, serotonine-creatinine-sulphate 100 micogram/rat, gamma-aminobutyric acid 100 microgram/rat, pilocarpine 1 mg/rat, histamine 100 microgram/rat were administered into the lateral ventricle of the brain. All agents were dissolved in Parker's fluid. Two control groups of animals were given Parker's fluid and subjected to surgical manipulations, respectively. Plasma TSH level was estimated after 30 min by means of radioimmunoassay. The increase in the TSH level was observed after the injection of serotonine and noradrenaline (4.0 and 3.1 ng/ml, respectively) as compared with control group (0.7 ng/ml).

The effect of serotonin given into the lateral ventricle of the brain on serum TSH level in normal and thyroxine-blocked rats.

The effect of serotonin given into the lateral ventricle of the brain in rats was investigated. Serotonin was solved in artificial cerebrospinal fluid and administered in three doses: 250 microgram, 50 microgram, 10 microgram. Only the highest dose of serotonin caused a statistically significant increase in serum TSH level 0.5 h after the injection. The possible role of serotonin in the blocking mechanism of thyroid hormones on TSH secretion was also taken into consideration. 50 microgram of serotonin significantly strengthened the lowering effect of 1-thyroxine given 0.5 h before on serum TSH level.