The dynamics of morphological changes during in vitro aging of bovine virgin mammary gland neutrophils

The present study was an in vitro analysis of the dynamics of bovine mammary gland neutrophil apoptosis based on the detection of morphological changes. The neutrophils were isolated from mammary glands of five virgin heifers. The mammary glands were lavaged, the suspensions were then bacteriologically examined, and total and differential cell counts were made. The cells were cultivated in vitro for 4 hours. After 2, 3 and 4 hours of cultivation, they were panoptically stained, and the proportions of apoptotic neutrophils and trypan blue positive neutrophils were determined. Neutrophil apoptosis and impaired cytoplasmic membrane integrity of neutrophils were already observed in the mammary gland lavages (11.9% and 0.8%, respectively). During the cultivation, a progressive increase in the number of apoptotic neutrophils in various stages of apoptosis – karyopyknosis, zeiosis and apoptotic bodies – was observed. Karyopyknotic neutrophils represented a dominant part of the apoptotic neutrophil population in the course of the whole cultivation. The most intensive increase was observed in zeiosis, whereas the levels of apoptotic bodies remained the same. After 4 hours of cultivation, 31.7% apoptotic neutrophils and 9.8% trypan blue positive neutrophils (i.e. Secondary necrotic cells) were found. The results of this work show that spontaneous apoptosis and secondary neutrophil necrosis must be taken into account during in vitro cultivations of bovine mammary gland neutrophils.

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Sladek Z, Rysanek D: CD14 expression, apoptosis and necrosis in resident and inflammatory macrophages from virgin bovine mammary gland

Veterinární Medicína ◽

10.17221/7777-vetmed ◽

2014 ◽

Vol 59 (No. 10) ◽

pp. 467-478 ◽

Cited By ~ 2

Author(s):

Z. Sladek ◽

D. Rysanek

Keyword(s):

Mammary Gland ◽

Mammary Glands ◽

Live Cells ◽

In Vitro Cultivation ◽

Bovine Mammary Gland ◽

Blood Monocytes ◽

Inflammatory Macrophages ◽

Apoptosis And Necrosis ◽

Phosphate Buffered Saline

This paper investigates the association between expression of CD14 and occurrence of apoptosis in blood monocytes, resident (RESMAC) and inflammatory macrophages from heifer mammary glands after infusion of PBS (INFMACPBS) or LPS (INFMACLPS). Resident macrophages (RESMAC) were obtained before, and inflammatory macrophages (INFMACPBS and INFMACLPS) 24 h after, induction of an inflammatory response using phosphate buffered saline (PBS) and lipopolysaccharide (LPS) in mammary glands of unbred heifers. Cell samples were analysed for differential counts, CD14 expression, apoptosis and necrosis using flow cytometry. In vitro cultivation led to a decrease in the proportion of living cells and to an increase in the proportion of apoptotic and necrotic cells in all macrophages and blood monocytes. In CD14+ macrophages, the proportions of live cells increased and proportions of apoptotic and necrotic cells decreased after in vitro cultivation. We observed in CD14− macrophages and monocytes that the proportions of live cells decreased and proportions of apoptotic and necrotic cells increased after in vitro cultivation. Our experiments confirm that the expression of CD14 in bovine mammary gland macrophages and blood monocytes is associated with cell viability.

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THE IN VITRO METABOLISM OF MAMMARY GLAND SLICES IN THE PRESENCE OF POSTERIOR PITUITARY LOBE EXTRACTS RICH IN MELANOPHORE-DISPERSING ACTIVITY

Journal of Endocrinology ◽

10.1677/joe.0.0150366 ◽

1957 ◽

Vol 15 (4) ◽

pp. 366-373 ◽

Cited By ~ 1

Author(s):

T. R. BRADLEY ◽

G. M. MITCHELL

Keyword(s):

Mammary Gland ◽

Guinea Pigs ◽

Incubation Medium ◽

Mammary Glands ◽

Gas Evolution ◽

Early Lactation ◽

Posterior Pituitary ◽

Rats And Mice ◽

Posterior Pituitary Lobe

SUMMARY Slices cut from mammary glands of rats and mice during gestation and lactation were incubated in vitro in the presence of pig posterior pituitary lobe extracts rich in melanophore-dispersing ('B') activity. Slices taken in early lactation but not during gestation or late lactation showed increased net gas evolution compared with control slices. Similar tissue from rabbits and guinea-pigs did not give rise to this effect, nor did slices of other tissues taken from lactating rats. The increased net gas evolution was not observed in the absence of glucose from the incubation medium. Treatment of the 'B' extract with NaOH or hypophysectomy of the rats prior to use decreased the response.

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Peer Review #3 of "TGF-beta signalling in bovine mammary gland involution and a comparative assessment of MAC-T and BME-UV1 cells as in vitro models for its study (v0.1)"

10.7287/peerj.6210v0.1/reviews/3 ◽

2019 ◽

Keyword(s):

Mammary Gland ◽

Peer Review ◽

In Vitro Models ◽

Comparative Assessment ◽

Bovine Mammary Gland ◽

Tgf Beta ◽

Mammary Gland Involution

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Evidence that the stimulation of lipogenesis in the mammary glands of starved lactating rats re-fed with a chow diet is dependent on continued hepatic gluconeogenesis during the absorptive period. Effects of a gluconeogenic inhibitory, mercaptopicolinic acid, in vivo

Biochemical Journal ◽

10.1042/bj2280727 ◽

1985 ◽

Vol 228 (3) ◽

pp. 727-733 ◽

Cited By ~ 8

Author(s):

D H Williamson ◽

V Ilic ◽

R G Jones

Keyword(s):

Mammary Gland ◽

Phosphoenolpyruvate Carboxykinase ◽

Mammary Glands ◽

Hepatic Glycogen ◽

Chow Diet ◽

Hepatic Gluconeogenesis ◽

Rapid Stimulation ◽

Stimulation Of

The rapid stimulation of lipogenesis in mammary gland that occurs on re-feeding starved lactating rats with a chow diet was decreased (60%) by injection of mercaptopicolinic acid, an inhibitor of hepatic gluconeogenesis at the phosphoenolpyruvate carboxykinase step. Mercaptopicolinate had no effect on lipogenesis in mammary glands of fed lactating rats. The inhibition of lipogenesis persisted in vitro when acini from mammary glands of re-fed rats treated with mercaptopicolinate were incubated with [1-14C]glucose. Mercaptopicolinate added in vitro had no significant effect on lipogenesis in acini from starved-re-fed lactating rats. Mercaptopicolinate prevented the deposition of glycogen and increased the rate of lipogenesis in livers of starved-re-fed lactating rats, whereas it had no significant effect on livers of fed lactating rats. Administration of intraperitoneal glucose restored the rate of mammary-gland lipogenesis in re-fed rats treated with mercaptopicolinate to the values for re-fed rats. Hepatic glycogen deposition was also restored, and the rate of hepatic lipogenesis was stimulated 5-fold. It is concluded that stimulation of mammary-gland lipogenesis on re-feeding with a chow diet after a period of starvation is in part dependent on continued hepatic gluconeogenesis during the absorptive period. Possible sources of the glucose precursors are discussed.

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