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Biomedical subjects

A H Pullen

Publications and source records attributed to A H Pullen.

27 records · Page 2Linked to original sources

Target dependence of Nissl body ultrastructure in cat thoracic motoneurones.

The effects on Nissl body (NB) ultrastructure of muscle reinnervation or neuroma formation were determined in cytochemically identified cat thoracic motoneurones subjected to axotomy by either nerve crush or nerve section with proximal ligation. Normal NB ultrastructure comprised highly ordered lamellae of rough endoplasmic reticulum (RER) with associated linear arrays of unbound polyribosomes. This ultrastructural orderliness was lost following axotomy, with or without light microscopic chromatolysis. While NBs were seen in the light microscope at late stages following both nerve crush or ligation, normal NB ultrastructure was only observed following nerve crush. An inductive effect of the periphery on NB ultrastructure is proposed and the implication of NB ultrastructure discussed in relation to protein synthesis.

Animals↗

A structured program in BASIC for cell morphometry: its application to the spinal motoneurone.

The BASIC program developed for use on an RML380Z computer collects, sorts, corrects and analyzes counts of organelle number, and measurements of membrane length and profile area. These fundamental parameters are used to derive intermediate parameters and their interrelationships are examined. Data input routines permit either measurements obtained from a digitizer tablet or the use of non-automated methods. A main program controls overall program flow and discrete sub-programs implement data input, data checking, derivation of parameters and analyses. The program is applied here to analyses of motoneurone synaptology, but with simple adaptation may be used to analyze other cell types.

Animals↗

Trophism between C-type axon terminals and thoracic motoneurones in the cat.

1. Quantitative ultrastructural examinations of axon terminals synapsing with normal alpha-motoneurones in segment T9 of cat spinal cord provided estimates of their numbers, sizes and synaptic structure. One synapse, the C type, derived from short-axon propriospinal segmental interneurones, was studied in detail.2. The numbers, sizes and post-synaptic structure of normal C-type synapses at T9 were compared with similar estimates from material provided by cats subjected to partial central deafferentation by double spinal hemisection at T5 and T10 between 7 days and 2 years previously.3. The proportion of C-type synapses present progessively increased from 1% in normal cats to 8.8% 200 days following hemisection, and had still attained a level of 3.1% by 2 years; these increases imply that the absolute number of C-type synapses underwent increase.4. Mean sizes of C-type synapses increased from 4.0 mum (normal) to 5.8 mum (200 days) and retained their enlarged sizes up to 2 years (5.9 mum). Furthermore, while 84% of C-type synapses were under 6 mum in length in normal motoneurones, 48% were over 6 mum long 200 days post-operatively.5. The unique post-synaptic structure of C-type synapses also proliferated following partial central deafferentation of the motoneurones. The elongated cistern, increased numbers and individual lengths of lamellae of the associated underlying rough endoplasmic reticulum indicated a trophic interaction between the presynaptic C terminal and its post-synaptic motoneurone.6. Counts of ribosomes ;bound' to lamellae of the subsynaptic rough endoplasmic reticulum, and of the lamellae-associated polyribosomes interposed between individual lamellae for normal and 200 day post-operative C-type synapses indicated an over-all post-operative increase in capacity for local subsynaptic protein synthesis topographically directed towards this type of axon terminal.7. The observed greater increase in frequency of ribosomes ;bound' to the rough endoplasmic reticulum, together with an over-all proliferation of this structure, specificially indicated an increased capacity for synthesis of protein for utilization in sites remote from those of synthesis (e.g. a trans-synaptic passage of protein).8. A hypothesis is advanced on the basis of the above results relating both pre- and post-synaptic changes in structure to an increased functional activation of the segmental short-axon propriospinal interneurones forming the C-type synapses, as a compensatory response to partial central deafferentation of spinal motoneurones.

Afferent Pathways↗

A structured program in BASIC for the analysis of peripheral nerve morphometry.

The BASIC program developed for use on a M.S.I. 6800 microprocessor analyses cross-sectional measurements of fibre (D) and axon (d) calibre made on electron micrographs of suitable magnification. Data input routines cover measurements of area and diameter, made manually or presented as fully computed values printed out by a M.O.P. Separate sub-routines input digitized data from a digitizer tablet coupled to the microprocessor. Myelin sheath thickness (2M) (D-d), or area and 'g' (d/D) are derived. The two analytical programs provide either frequency distributions, with associated statistics for each parameter, or analyses of the relationships (D:d), (d:2M) and (D:g). The highly structured nature of the program permits it to be edited or extended according to the project. The main programme controls the overall flow of analysis; discrete subprograms inserted into the main program perform the data input, parameter derivations, data checks and analyses. This system offers maximum flexibility and is suitable for most morphometric investigations involving peripheral nerves.

Animals↗

A quantitative ultrastructural study of dorsal root regeneration.

Regeneration in rat lumbo-sacral dorsal roots was studied 5-71 days following crush lesions. Wallerian degeneration occurred up to 20 days. At 11 days degenerating myelin was found in both Schwann cells and macrophages. Myelination was first observed 4 mm central to the crush at 7 days, and myelin became compact when the mesaxon exhibited 3.5 turns about the axon (about 11 days post-operatively). At 71 days, 69% of all fibres were myelinated, compared with 36% in normal roots. An example of 2 axons myelinating within the same Schwann cell occurred at 20 days. In normal roots curvilinear relationships were found between axon diameter and fibre diameter, myelin thickness and axon diameter, and between g and fibre diameter. In contrast, linear relationships between these parameters occurred in post-operative roots up to 71 days. Curvilinearity returned at 71 days. Alterations in the relationship between axon diameter and myelin thickness during regeneration indicated that myelin growth lagged behind axon growth throughout, but was more noticeable in larger calibre fibres. By 71 days, larger fibres exhibited disproportionately thin myelin, whilst small fibres possessed abnormally thick myelin compared to normal fibres of similar calibre. Regeneration was limited by axons failing to make successful central synaptic connections and by the poor metabolic response of dorsal root ganglion cells to sectioning of their central processes.

Animals↗

The distribution and relative sized of fibre types in the extensor digitorum longus and soleus muscles of the adult rat.

Histochemical fibre types classified in sections stained for succinic dehydrogenase (sdh) and myosin ATPase at pH 9.4, were found to be distributed in a consistent manner within the extensor digitorum longus (EDL) and 4 soleus muscles of the adult rat. Simple morphometric techniques applied to complete transverse sections of both muscles showed that the relative distributions and proportions of fibre types along their deep to superficial, and medial to lateral, axes varied accoording to the histochemical method used for fibre typing. Similar differences occurred when the relative ranges of size exhibited by each fibre type were compared in sections stained for SDH and ATPase, and the discrepancies in fibre classification were confirmed by an analysis of individual fibres in serial sections. The findings are discussed in relation to those previosly reported for the EDL and soleus muscles of the rat.

Adenosine Triphosphatases↗

The distribution and relative sizes of three histochemical fibre types in the rat tibialis anterior muscle.

A quantitative study of the tibialis anterior muscle of the adult rat showed that the proportions of each 'histochemical' fibre type varied between adjacent regions along the deep to superficial and medial to lateral axes of the muscle. The distribution of fibre types classified with oxidative enzymes was similar to that using phosphorylase, but differed from that obtained with sections stained for myosin ATPase. This apparent discrepancy in classifation was confirmed by comparison of the ranges of fibre cross sectional areas for each fibre type classified on the basis of their oxidative enzme and ATPase activites, and by an independent analysis of individual fibres in serial sections stained for succinic dehydrogenase and ATPase.

Adenosine Triphosphatases↗

A parametric analysis of the growing CFHB (Wistar) rat.

Measurements of body weight and body, tail and hindlimb lengths were made at various times during the postnatal development of the CFHB-Wistar rat using simple techniques which avoided unnecessary stress being applied to the animal. All the graphical relationships studied using these measurements indicated that growth was divided into two phases which meet at 16 days. This was particularly noticeable when either body length or hindlimb length was plotted against either age or body weight. Body weight appeared to be a better indication of maturity than time; and linear relationships were found when measurements of body lengths were plotted against the square root of body weight. Curvilinear graphs were obtained when body weight was plotted on alternative scales. Results suggest that prior to 16 days metabolism is probably directed more towards attaining sufficient maturity to enable the animal to survive independently, than to increasing its stature; but after 16 days this situation is reversed.

Age Factors↗