A small scale cell culture system to analyze mechanobiology using reporter gene constructs and polyurethane dishes.

Mechanical forces are translated into biochemical signals and contribute to cell differentiation and phenotype maintenance. Mesenchymal stem cells and their tissue-specific offspring, as osteoblasts and chondrocytes, cells of cardiovascular tissues and lung cells are sensitive to mechanical loading but molecules and mechanisms involved have to be unraveled. It is well established that cellular mechanotransduction is mediated e.g. by activation of the transcription factor SP1 and by kinase signaling cascades resulting in the activation of the AP1 complex. To investigate cellular mechanisms involved in mechanotransduction and to analyze substances, which modulate cellular mechanosensitivity reporter gene constructs, which can be transfected into cells of interest might be helpful. Suitable small-scale bioreactor systems and mechanosensitive reporter gene constructs are lacking. To analyze the molecular mechanisms of mechanotransduction and its crosstalk with biochemically induced signal transduction, AP1 and SP1 luciferase reporter gene constructs were cloned and transfected into various cell lines and primary cells. A newly developed bioreactor and small-scale 24-well polyurethane dishes were used to apply cyclic stretching to the transfected cells. 1 Hz cyclic stretching for 30 min in this system resulted in a significant stimulation of AP1 and SP1 mediated luciferase activity compared to unstimulated cells. In summary we describe a small-scale cell culture/bioreactor system capable of analyzing subcellular crosstalk mechanisms in mechanotransduction, mechanosensitivity of primary cells and of screening the activity of putative mechanosensitizers as new targets, e.g. for the treatment of bone loss caused by both disuse and signal transduction related alterations of mechanotransduction.

Mother is not like mother: Concurrent pregnancy reduces lactating guinea pigs' responsiveness to pup calls.

Offspring signalling can serve to communicate need to the parents and thus influence parental readiness to provide care. Offspring stimuli that affect parental care have been investigated extensively. Yet much less is known about the mechanisms leading to a decline in maternal motivation when conflicts of provisioning current and future offspring may arise. Here we tested responses by pregnant and non-pregnant female guinea pigs (Cavia aperea f. porcellus) to playback of pup calls during their period of lactation for dependent offspring. Most concurrently pregnant and lactating females did not respond to pup calls, whereas non-pregnant lactating females responded strongly. Our findings expand on previous studies by showing that female behavioural responsiveness to pup stimuli is strongly reduced by concurrent pregnancy and lactation. These instantaneous measurements of female responsiveness to young show more directly than standard measures like nursing performance or time to weaning how female motivation to care for current offspring is diminished by simultaneous gestation.

Vocal mother-offspring communication in guinea pigs: females adjust maternal responsiveness to litter size.

BACKGROUND: In parent-offspring communication, vocal signals are often used to attract attention and offspring might call to induce parental behaviour. In guinea pigs (Cavia aperea f. porcellus) mothers wean larger litters later than small ones, but it is unknown whether this difference depends on processes induced during pregnancy or is influenced post-natally by the number of pups present. We here tested with playback-experiments using pup separation calls whether mothers with cross-fostered large experimental litters (four-pup-litters) were more responsive to offspring calls and maintained responsiveness for longer than mothers with small experimental litters (two-pup-litters). Mothers were tested when two pups were suckling i.e. when both teats were occupied. RESULTS: Mothers of four-pup litters responded stronger to broadcast pup separation calls than those with two-pup litters. Additionally, we tested the mothers' responsiveness to pup separation calls in the absence of their pups on day 8 and 20 of lactation. Mothers of four-pup litters responded stronger and showed no decrease in responsiveness from day 8 to 20, whereas mothers of two-pup litters responded less and decreased responsiveness from day 8 to 20. Mothers of four-pup litters also weaned their pups 5 days later than those of two-pup litters. CONCLUSION: Measured by their response to pup calls and by time to weaning, guinea pig mothers adjust maternal responsiveness to litter size. This behaviour is likely to be an adaptive strategy in resource allocation during reproduction.