PubMed Health⌕ Search

Biomedical subjects

L J McGuffee

Publications and source records attributed to L J McGuffee.

25 records · Page 2Linked to original sources

A 45Ca autoradiographic and stereological study of freeze-dried smooth muscle of the guinea pig vas deferens.

In an effort to more clearly elucidate the role of cellular structures as calcium sinks and sources in smooth muscle cells, the intracellular distribution of radioactive calcium was evaluated by a new method based on freeze-drying. The guinea pig vas deferens was exposed to a physiological salt solution that contained 45Ca. The muscle was then freeze-dried and prepared for electron microscope autoradiography. The grain density over the plasma membrane, mitochondria, and sarcoplasmic reticulum (SR) was significantly greater than that of the matrix. These results suggest that the plasma membrane, mitochondria and SR have the capacity to accumulate calcium. Which of these structures serve as a source of calcium for contraction remains to be determined. A stereological comparison between freeze-dried and conventionally prepared smooth muscles revealed several differences. The cross-sectional area of freeze-dried cells was about twice that of conventionally prepared cells. Moreover, mitochondria and sub-surface vesicles occupied a significantly smaller percentage of the cell in the freeze-dried tissue than they did in the conventionally prepared tissue.

Animals↗

Ultrastructural characteristics of freeze-dried smooth muscle.

The rapid freezing and vacuum dehydration of tissue has been employed to study the intracellular distribution of diffusible substances at the electron microscopic level. Hower, the ultrastructural detail of freeze-dried tissue is difficult to retain. Consequently, the ultrastructural preservation of freeze-dried smooth muscle has not been sufficient to permit satisfactorily definition of intracellular organelles. Therefore, determinations of the intracellular distribution of soluble ions have not been achieved in freeze-dried smooth muscle. In this study a freeze drying method is presented which provides more satisfactory definition of intracellular organelles than has been available in the past. Using this merens have been preserved. When these muscles are compared to convention preparer of surface vesicles that are observed, and the mitochondria are extremely electron opaque in the freeze-dried tissue. The smooth muscles which have been examined do not have the inherent contrast of other types of tissue nor do they contain the different types of mitochondria that have been observed in nonmuscle tissue.

Animals↗

Ultrastructure, calcium accumulation, and contractile response in smooth muscle.

After removing extracellular Ca2+ with [ethylene bis(oxyethylenenitrilo)]tetraacetic acid, we found that the guinea pig vas deferens (VD) was mechanically responsive to electrical stimulation for a significantly greater length of time than was guinea pig taenia coli (TC). An obvious explanation for these findings is that the VD has more intracellular calcium available for contraction than does the TC. To determine if this explanation is plausible, the volume of internal storage structures within the two muscles was compared. It was found that the volumes of potential sequestering structures in the VD and TC are not significantly different. Next, the affinities of the storage structures for calcium were compared. The VD was found to accumulate approximately twice as much 45Ca as did the TC, as determined by 45Ca autoradiography. Calcium-45 was present to a greater extent in association with surface vesicles, sarcoplasmic reticulum (SR), and mitochondria than in the unassociated state within the cytoplasmic matrix. Based on the results of these experiments, we suggest that the VD and the TC of the guinea pig differ in the affinity of their storage sites for calcium.

Animals↗