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R B Borgens

Publications and source records attributed to R B Borgens.

49 records · Page 3Linked to original sources

Stump currents in regenerating salamanders and newts.

We report here that a variety of salamanders and newts from differing habitats all drive a steady ionic electric current out of the forelimb stump tip after forelimb amputation. Several hours after amputation the density of this stump current ranges from about 10 to 100 microA/cm2 in most species, and declines with time. In most cases, the magnitude of the stump current is dependent on the concentration of Na+ in the external medium (an artificial pondwater), suggesting that the well-known Na+ -dependent transcutaneous voltage described in amphibia (particularly frogs) is the EMF for this stump current. These measurements add to those previously reported for the North American red spotted newt (Notophthalmus viridescens), and suggest that electrical changes following amputation of urodele limbs are widespread among members of this group.

Amputation, Surgical↗

Are limb development and limb regeneration both initiated by an integumentary wounding? A hypothesis.

It is proposed that, whereas an actual wound to a salamander limb may initiate limb regeneration, a local and developmentally programmed integumentary wound may initiate limb development. The electrophysiological changes induced by these lesions of the skin may be a common denominator linking limb regeneration and limb development. Such early electrical events are considered to initiate or guide the early accumulation of cells, and to help to produce the local environment in which a limb will arise. This scheme provides a self-limiting positive-feedback mechanism for the production of a localized area where other developmental mechanisms act in concert with endogenous electrical fields (or in their complete absence), thereby leading to limb differentiation. This hypothesis may not be restricted to limb formation; it may be of more general significance, i.e. in the process of organogenesis in embryos. One might reasonably suggest that, by such a mechanism, any developing placode (for example, auditory or olfactory placodes) might form and localize.

Amphibians↗

A steady efflux of ionic current predicts hind limb development in the axolotl.

For more than a week prior to the emergence of a hind limb, a steady electric current is driven out of the ventrolateral flank in the immature axolotl, returning through the integument in adjacent regions of the body. A marked peak in the density of this outcurrent could be observed over the exact area of hind limb formation 4 to 6 days prior to its appearance. After a bud projected from the flank, current densities were observed to decrease in magnitude yet localize about the early limb. In about one-half of the animals observed, current reversed its polarity and entered the apex of large buds, 0.4 to 0.5 mm in length. We discuss the possible role such endogenous currents and their associated fields may play in limb development and compare it to similar current flow associated with the regeneration of amphibian limbs.

Ambystoma↗

Mice regrow the tips of their foretoes.

Mice will replace the tip of a foretoe when it is amputated distal to the last interphalangeal joint. Amputation of the digit more proximal to the joint does not result in regrowth of the foretoe. Though this growth shares certain similarities with the epimorphic regeneration of amphibian limbs, the two processes are not the same. The regrowth reported here in mice is probably similar to the scattered clinical reports of fingertips regeneration in children, and presents a model system with which to explore the controls of wound healing and tissue reconstruction in mammals.

Amputation, Surgical↗

Enhanced spinal cord regeneration in lamprey by applied electric fields.

After a weak, steady electric current of approximately 10 microamperes was imposed across the completely severed spinal cord of the larval lamprey Petromyzon marinus, enhanced regeneration was observed in the severed giant reticulospinal neurons. The current was applied with implanted wick electrodes for 5 to 6 days after transection (cathode distal to lesion). The spinal cords were examined 44 to 63 days after the operation by means of intracellular fluorescent dye injections and electrophysiology. Extracellular stimulation of whole cords showed that action potentials in most of the electrically treated preparations were conducted in both directions across the lesion, but they were not conducted in either direction in most of the sham-treated controls. In most of the electrically treated animals, processes from giant axons with swollen irregular tips, indicating active growth, were seen in or across the lesion. Only a few of the sham-treated controls showed these features. It is possible that these facilitated regenerative responses were mediated by the effects of the artificially applied electric fields on the natural steady current of injury entering the spinal lesion.

Action Potentials↗

Large and persistent electrical currents enter the transected lamprey spinal cord.

The electrical currents at the surface of the proximal portion of an isolated and transected lamprey spinal cord were measured with an extracellular vibrating probe. Soon after transection, currents of about 0.5 mA/cm2 enter the cut surface of the spinal cord. These currents fall to about a quarter of their initial value within an hour; within the next 2 days they gradually decline from about 100 microA/cm2 to about 4 microA/cm2; they then remain constant up to 6 days posttransection, when the measurements were ended. The pattern of current entry included substantial peaks opposite (and presumably into) the cut ends of giant axons. Response to changes in the ionic composition of the medium indicates that about half of the injury current consists of Na+, and that much of the rest may consist of Ca2+. The measured influx of ions, which adds up to several coulombs per cm2 in a few days, should radically alter the ionic composition of the terminal few millimeters of neural tissue. Thus it may be important in the degenerative and regenerative responses of neurons to axotomy.

Action Potentials↗

Role of subdermal current shunts in the failure of frogs to regenerate.

Large, uniform, skin-driven currents (20-40 muamp/cm2) leave the ends of limb stumps of post-metamorphic frogs (Rana pipiens) from about the first through the tenth day after amputation. However, right after amputation, while currents of comparable density may leave the periphery of the cut surface, current densities are greatly depressed in the center of this surface. We suggest that this depression is brought about by shunting through the subdermal lymph space (characteristic of anurans but not urodeles); continues in covert form after formation of a wound epithelium; and helps explain the ability of small, imposed currents to initiate frog limb regeneration.

Action Potentials↗

Reduction of sodium dependent stump currents disturbs urodele limb regeneration.

We have asked the question whether the natural electric currents which leave urodele limb stumps are in any way needed for their regeneration. As an initial test, we have greatly reduced such currents in the tiger salamander, Ambystoma tigrinum, by applying 0.5 mM amiloride to the stump skin or by immersion of the animals in sodium depleted media. We have also reduced such currents in the red spotted newt, Notophthalmus viridescens, by such immersion. Limb regeneration in half of the amiloride-treated animals was either entirely blocked or grossly deficient, while the others regenerated normally. Limb regeneration in sodium depleted media was consistently inhibited for some weeks but then recovered. These results are consistent with the hypothesis that stump currents are in some way needed for normal regeneration.

Ambystoma↗

Bioelectricity and regeneration: large currents leave the stumps of regenerating newt limbs.

Electrical currents near regenerating newt limbs were measured with a recently developed vibrating probe. Steady currents with local surface densities of 10 to 100 muA/cm2 or more leave the end of the stump during the first 5-10 days after amputation and are balanced by currents with densities of only 1-3 muA/cm2 that enter the intact skin around the stump. They are immediately dependent upon the entry of sodium ions into this skin and are therefore inferred to be skin-driven. The outward currents are comparable in direction, density, duration, and position to artificially imposed currents previously found sufficient to induce significant regeneration of amputated adult frog limbs. This comparison suggests that the endogenous stump currents play some causal role in initiating regeneration.

Amiloride↗

Advances in three-dimensional reconstruction of the experimental spinal cord injury.

Three-dimensional (3D) computer reconstruction is an ideal tool for evaluating the centralized pathology of mammalian spinal cord injury (SCI) where multiple anatomical features are embedded within each other. Here, we evaluate three different reconstruction algorithms to three-dimensionally visualize SCIs. We also show for the first time, that determination of the volume and surface area of pathological features is possible using the reconstructed 3D images themselves. We compare these measurements to those calculated by older morphometric approaches. Finally, we demonstrate dynamic navigation into a 3D spinal cord reconstruction.

Animals↗